Qualitative Assessment of Bacteria and Fungi in the ...

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Pakistan Journal of Scientific and Industrial Research Series B: Biological Sciences Vol. 55, No. 3, November-December, 2012 Contents Effect of Foliar Applied Boron Application on Growth, Yield and Quality of Maize (Zea mays L.) Muhammad Tahir, Asghar Ali, Farhan Khalid, Muhammad Naeem, Naeem Fiaz and Muhammad Waseem

Diversity Analysis of Marigolds – Tagetes (Asteraceae)

Chithra Santhakumari Amma and Rajalakshmi Radhakrishnan

A Study of Poisonous Plants of Islamabad Area, Pakistan Saleem Ahmad

Effects of Passiflora foetida Linn. (Passifloraceae) on Genital Tract, Serum Estradiol, Pituitary Gonadotropin and Prolactin Level in Female Adult and Immature Ovariectomized Rats Bleu Gome Michel, Kouakou Koffi, Toure Alassane and Traore Flavien

Growth Performance of Juvenile Milkfish Chanos chanos (Forskal) on Replacement of Fish Meal with Plant Based Diet Supplemented with Dietary Cell Bound Phytase of Pichia anomala Sajad Hassan, Karim Altaff , Tulsi Satyanarayana, Syed Ahamed Ali and Thirunavuarasu

Detoxification of Aflatoxin B1 in Poultry and Fish Feed by Various Chemicals Alim-un-Nisa, Naseem Zahra, Sajila Hina and Nusrat Ejaz

Qualitative Assessment of Bacteria and Fungi in the Indoor Environment of Hospitals of Islamabad, Pakistan Fouzia Hussain, Subhe Sadiq Tahir, Naseem Rauf and Afifa Batool

117 122 129

138

145 154

159

Short Communications Effects of Some Oral Hypoglycaemic Drugs on Erythrocyte Nicotinamide Adenine Dinucleotide Hydrogen Diaphorase (E.C.1.6.4.3) Activity of Wistar Albino Rats (Rattus rattus) Samuel Chimezie Onuoha and Austin Amadike Uwakwe

Estimation of Heavy Metals in Medicinal Plants as a Source of Herbal Medicine Used in Cardiovascular Diseases Farah Deeba, Naeem Abbas and Rauf Ahmed

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169

Assessment of Selected Quality Attributes of Jam Formulated from Baobab-Hogplum Fruits

Adekanmi Oyeyinka Abioye, Samson Adeoye Oyeyinka, Adewumi Toyin Oyeyinka and Sarafadeen Omowumi Kareem

172

Conetnts of Volume 55, Ser. B: Biol. Sci. (No. 1-3)

i

Author Index of Volume 55, Ser. B: Biol. Sci.

iv

Subject Index of Volume 55, Ser. A: Phys. Sci.

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Pak. j. sci. ind. res. Ser. B: biol. sci. 2012 55 (3) 117-121

Effect of Foliar Applied Boron Application on Growth, Yield and Quality of Maize (Zea mays L.) Muhammad Tahir a, Asghar Ali a, Farhan Khalid a, Muhammad Naeem b, Naeem Fiaz a and Muhammad Waseem a* a

b University

Department of Agronomy, University of Agriculture, Faisalabad, Pakistan College of Agriculture and Environmental Sciences, Islamia University, Bahawalpur, Pakistan (received April 7, 2011; revised December 17, 2011; accepted January 9, 2012)

Abstract. A field study was carried out to evaluate the effect of foliar applied boron application on growth, yield and quality of maize (Zea mays L.). Foliar application of boron was carried out after 20 days of crop emergence at 0, 0.15, 0.30 and 0.45 kg of B/ha. Boron application at 0.30 kg/ha increased the plant height, leaf area, stem diameter, cob weight, number of grains per cob, protein and oil contents. The maximum grain yield (7.14 tons/ha) and biological yield (527.4 tons/ha) was recorded in B2 where application of boron was carried out at 0.30 kg/ha, however, further increase in boron dose decreased the yields. Keywords: maize, hybrid, boron, foliar spray, cereal, micronutrient

Introduction

(Shorrocks, 1997). Deficiency symptoms of boron can be observed in vegetative and reproductive parts such as inhibition of growth of shoot and root tips, flower development, reduced setting and malformation of seeds. However, application of boron require thorough analysis of requirements as excessive application of boron is toxic for plant growth (Dell and Huang, 1997; Sherrell, 1983). The present study was carried out to investigate the influence of foliar applied boron concentrations on growth and yield of maize hybrid.

Cereals are important components of human diet providing 65% of the energy requirements. The cultivation and domestication of these crops has long standing history. Among cereal crops, maize is at the top in terms of its production share. However in Pakistan, it ranks third after wheat and rice. Presently it is grown approximately on 1015 thousand hectares with total production of 3313 thousand tons (NNS, 2008). Maize cultivation is beneficial as its processing yield several products that have the important industrial applications (Bressani et al., 2004) resulting in its increased demand. There are several factors that play dominant role in enhancing its yield; proper and balance fertilization is one of the important factors in increasing the yield and quality of the crop. The present scenario suggests that application of macronutrients is also essential along with macronutrients to get the desired results with respect to biological and grain yield. Moreover, Pakistani soils are deficient in boron, zinc etc. (Hussain et al., 2006).

Materials and Methods The field experiment was conducted at Agronomic Research Area, University of Agriculture, Faisalabad, during spring 2008 to evaluate the influence of foliar applied boron concentrations on growth and yield of maize (Zea mays L.) hybrid. A factorial experiment was organized in randomized complete block design (RCBD) with four replications having net plot size of 6 m ´ 3 m. Maize hybrid Monsanto-6525 was sown in 2nd week of March in 75 cm apart ridges in hills 30 cm apart at one side of the ridge using seed rate of 25 kg/ha. The fertilizers were applied at 250-125-125 kg/NPK/ha as Urea, DAP and SSP, respectively, dose of phosphorous and potash and half dose of nitrogen at sowing and remaining half nitrogen dose was applied at top dressed at knee height. Boron was applied at 0, 0.15, 0.30 and 0.45 kg/ha as foliar spray 20 days after crop emergence. Soil samples were taken before sowing of crop to a depth of 30 cm for physiochemical analysis. Soil was loamy type with pH of 7.87, EC of 2.16 dSm and SAR

Boron deficiency in crops is more widespread than that of the other micronutrients (Gupta et al., 1985). Boron is closely associated with the growth of plants and plays a vital role in cell division. Moreover, requirements of boron vary according to species, crops and stages i.e. reproduction and vegetative. The requirement of boron to crops is affected by several environmental factors like temperature, light and soil water conditions *Author for correspondence; E-mail: [email protected]

117

118

Muhammad Waseem et al.

440 ppm. Likewise, it contained organic matter (0.49%), phosphorous (4.08 ppm), potassium (273 ppm) and boron (0.32 mg/kg). This analysis showed that soil deficient in boron contents can not meets the requirements of maize crop. All other agronomic practices were kept normal and uniform for all the treatments. Growth and yield parameters. Plant height (cm), leaf area (cm2), stem diameter (cm), cob weight (g), cob length (cm), cob diameter (cm), number of grains per cob, 100 grain weight (g), grain rows per cob, biological yield (tons/ha), grain yield (tons/ha), protein contents (%) and oil contents (%) were recorded. Data collected were analyzed statistically using Fisher's analysis of variance technique. Difference among the treatment means was compared using least significant difference at 5 % probability level (Steel et al., 1997).

with an increasing trend with increasing concentrations of boron (Fig. 1). Both the higher and lower levels of boron where boron was applied @ 0.15 and 0.45 kg/ha, respectively, decreased the cob weight significantly when compared with boron applied @ 0.30 kg/ha. The cob length was not affected significantly by boron application and values were in the range of 19.97 to 21.57 cm. The relationship of cob diameter showed a slightly increasing trend increasing concentration of boron (Fig. 2). Maximum cob diameter was recorded in fields where foliar spray of boron @ 0.30 kg/ha was applied. However, differences among different boron levels were not significant.

170

y = 130.87 + 37.28 x R2 = 0.26

160

Indices of growth. The growth indices of maize indicated the significant influence of boron application. The increasing tendency in plant height, leaf area and stem diameter was observed from 153.4 to 163.1 cm, 15.82 to 19.00 cm2 and 1.867 to 3.138 cm, respectively, when boron was applied @ 0.30 kg/ha (Table 1). The maximum plant height, leaf area and stem diameter were recorded where boron was applied @ 0.30 kg/ha with mean values of 163.1 cm, 19.0 cm2 and 3.138 cm, respectively. However, increasing the dose of boron from 0.30 to 0.45 kg/ha, resulted in significant declining in all attributes of growth but was still significantly higher as compared to control. Cob characteristics. Cob characteristics include cob weight (g), cob length (cm) and cob diameter (cm). These all parameters are indicator of better yield performance. The cob weight showed significant results

Cob weight (g)

Results and Discussion

150

140

130

120 0.00

0.15

0.30

0.45

Concentrations of boron (kg/ha)

Fig. 1. Effect of foliar applied boron concentrations on cob weight. Yield indices. The yield indices include number of grains per cob, 100 grain weight, grain rows per cob, biological yield and grain yield. Data concerning number

Table 1. Effect of foliar applied boron application on growth, yield and quality of maize at maturity stage Treatments

Plant height (cm)

Leaf area (cm2)

Stem diameter (cm)

Cob length (cm)

Grain rows/ cob

Biological Grain yield yield (tons/ha) (tons/ha)

Protein contents (%)

Oil contents (%)

B0 = Control B1 = 0.15 kg/ha B2 = 0.30 kg/ha B3 = 0.45 kg/ha F-value LSD value

153.4c 159.1b 163.1a 157.4b 18.7704** 2.978

15.82d 16.77c 19.00a 17.65b 33.594** 0.7451

1.867c 2.395b 3.138a 2.557b 55.2245** 0.2262

19.97 21.27 21.53 21.57 1.138ns --

14.50 15.50 15.54 14.32 0.603ns --

333.1c 430.4b 527.4a 426.8b 37.305** 41.54

9.50c 10.62b 11.23a 10.57b 30.6768** 0.4140

4.39c 4.71b 4.84a 4.35c 53.0804** 0.1012

6.045d 6.883b 7.145a 6.523c 87.0512** 0.160

Mean values in column not comprising the same letter (a, b, c, d) vary significantly at (P=0.05); ** = highly significant at 5% level; ns = non significant.

119

Foliar Applied Boron for Maize Growth 5.00

sively and hold linear relationship with born application (Fig. 4). The increased boron dose from 1.5 to 0.45 kg/ha significantly decreased the 100 grain weight which, however was statistically similar to the 0.15 kg/ha. The minimum 100 grain weight was recorded in control treatment. In contrary, number of grain rows per cob was not significantly affected due to foliar application of boron at varying doses (Table 1).

y = 3.97 + 0.05 x R 2 = 0.37

Cob diameter (cm)

4.50

4.00

3.50

3.00 0

0.15

0.3

0.45

Concentrations of boron (kg/ha)

Fig. 2. Effect of foliar applied boron concentrations on cob diameter. of grains per cob showed highly significant results and its linear relationship showed an increasing trend with different boron concentrations (Fig. 3). Relationship showed that with the increase in boron concentration there was an increase in number of grains per cob up to a certain level while its excessive dose lowered the number of grains. Application of boron significantly increased the number of grains per cob compared to control. The difference among different boron levels could not reach to the level of significance. The data pertaining to 100 grain weight when subjected to statistical analysis revealed significant differences among treatments. The attribute increased progres-

As far as biological and grain yields are concerned, foliar application of born improved the biological yield significantly. The lowest biological yield of 333.1 tons/ha was recorded in control fields as compared to maximum (527.4 tons/ha) in maize subjected to foliar application of boron @ 0.30 kg/ha. Moreover, boron @ 0.15 and 0.45 kg/ha were statistically at par with mean yield of 430.4 and 426.8 tons/ha, respectively. Foliar application of boron showed highly significant results for grain yield. Comparison of mean values (Table 1) showed that maximum grain yield of 7.14 tons/ha was recorded when boron was applied @ 0.30 kg/ha followed by boron applied @ 0.15 kg/ha (6.88 tons/ha). The increase in boron level from 1.5 to 0.45 kg/ha resulted in sharp reduction in grain yield and grain yield obtained from B3 (6.52 tons/ha) was significantly lower from B2 (7.145 tons/ha) and B1 (6.88 tons/ha) while the least grain yield was recorded in control (6.04 tons/ha) where boron was not applied. Qualitative traits. Boron application significantly affected protein contents of grains and maximum protein 40.00

y = 503.93 + 9.15 x R2 = 0.68

550.00

35.00 100-grain weight (g)

Number of grains/cob

540.00

530.00

520.00

30.00

25.00

510.00

500.00

y = 25.36 + 2.08 x R 2 = 0.38

0

20.00 0.15

0.3

0.45

Concentrations of boron (kg/ha)

Fig. 3. Effect of foliar applied boron concentrations on number of grains per cob.

0

0.15

0.3

0.45

Concentrations of boron (kg/ha)

Fig. 4. Effect of foliar applied boron concentrations on 100-grains weight.

120 contents (11.23 %) were recorded where boron was applied @ 0.30 kg/ha while higher and lower doses, than this, were found to be similar with mean oil contents of 10.62 and 10.57 %, but were higher than control (9.50 %) where boron application was not carried out (Table 1). Oil contents is an important factor that contributes towards the yield and boron application at various rates showed momentous effect on oil contents (Table 1). Boron application @ 0.30 kg/ha showed maximum oil contents (4.84%) followed by B2 (4.71%) where boron was applied @ 0.15 kg/ha. While higher dose of 0.45 kg/ha showed similar results to control with oil contents of 4.35 and 4.39, respectively. Boron has been found to be an essential micronutrient element for plant growth and development. Previous studies showed that boron is a micronutrient that is essential for plant only but recent research work carried out by different research workers had shown that boron is essential for animals especially human beings because of its key role in synthesis of estrogen, vitamin D and other steroid hormones. Indices of growth. In this study, increasing concentration of boron significantly affected the growth parameter like plant height, leaf area and stem diameter. Momentous increase recorded in growth indices of maize plants might be due to its vital role in cell wall synthesis, division, elongation and nucleic acid metabolism. Reduction in mean values recorded for growth parameters at higher dosage of boron application could be due to the burning effect of boron noted on leaves. Findings of present research are well in agreement with that of Ceyhan et al. (2007) and Mazher et al. (2006). They are of the view that boron application improves the growth parameters significantly. Moreover, Ahmed et al. (2008) also highlighted the role of boron applied at various rates and found significant improvements in growth indices. Similar results were also reported by Lopez-Lefebre et al. (2002) who reported marked differences among the means regarding biomass production. Cob characteristics. Cob characteristics include cob weight, cob length, cob diameter and number of gains per cob and these parameters contribute towards the final yield. The improvements in cob characteristics of maize hybrid bolster the prospects of boron applications for the enhancement of growth and maize yield. The momentous effect of boron application was found

Muhammad Waseem et al.

regarding the cob related features of the maize plants. This significant effect of boron application might be due to its vital role in pollen tube formation, increasing the efficiency of fertilization process, which may results in variability among the mean values regarding cob weight, cob diameter and number of grains per cob. This may result in improvement of the grain size of the plants. These results are in accordance with Gupta (1993), who reported that boron is required continuously for growth of most of the plants otherwise deficiency occurs. The reduction in cob weight at higher dose may be due to the toxic effects of boron leading to burning of leaf margins and affecting adversely the photosynthetic activity of plant while in control reduction in cob weight may be due to the deficiency of boron. The results indicated that boron has significant effect on number of grains per cob while in case of deficiency the number of grains per cob may reduce. Yield indices. The yield indices include 100 grain weight, grain rows per cob, biological yield and grain yield. The improvement in grain and biological yield of maize hybrids is mainly attributed to complementary role of boron in the reproduction and vegetative stage of plants. Usually, micronutrients are required in minute amounts but impart significant effects on metabolism by working synergistically with hormones and enzymes in normal functionality of system. The present findings are well in agreement with that of Ceyhan et al. (2008), Blamey et al. (1997) and Schon and Blevins (1990). They all are of the view that boron application is necessary in all those soils which are deficient in micronutrients. Moreover, Donald et al. (1998) suggested the foliar application as best mode of fertilizer application due to its better efficiency. Qualitative traits. Protein and oil contents mainly contribute towards the yield of the plants and are affected by application of different micronutrients including boron. The decrease in protein and oil contents at higher dose might be due to burning of the leaves that affected its photosynthetic performance and other physiological functions. Boron has been found to play a key role in nitrogen metabolism and assimilation (Ruiz et al., 1998; Shelp, 1988). Nitrogen is a primary constituent of protein as well as all enzymes (Raun and Johnson, 1999) and in this way boron application showed marked influence on protein and oil contents of grains.

Foliar Applied Boron for Maize Growth

Conclusion Present study highlighted the importance of boron application in soils which are boron deficient especially in Pakistan. Boron application nearly improved all the traits at dosage of 0.15 and 0.30 kg/ha, however, at higher dosage of 0.45 kg/ha declining tendency was observed due to its toxic effect and caused burning of the leaves. Overall, boron @ 0.30 kg/ha is recommended due to its role in improving growth and yield indices.

References Ahmed, N., Abid, M., Ahmad, F. 2008. Boron toxicity in irrigated cotton (Gossypium hirsutum L.). Pakistan Journal of Botany, 40: 2443-2452. Blamey, F.P.C., Zollinger, R.K., Schneiter, A.A. 1997. Sunflower production and culture; In: Sunflower Science and Tehnology. A. A. Schneiter (ed.), pp. 595-670, Agronomy Monograph 35, ASA, CSSA and SSSA Madison, WI, USA. Bressani, R., Turcios, J.C., Ruiz, A.S.C., Palomo, P.P. 2004. Effect of processing conditions on phytic acid, calcium, iron, and zinc contents of lime-cooked maize. Journal of Agricultural Food Chemistry, 52: 1157-1162. Ceyhan, E., Onder, M., Ozturk, O., Harmankaya, M., Hamurcu, M., Gezgin, S. 2008. Effects of application boron on yields, yield component and oil content of sunflower in boron-deficient calcareous soils. African Journal of Biotechnology, 7: 2854-2861. Ceyhan, E., Onder, M., Harmankaya, M., Gezgin, S. 2007. Response of chickpea cultivars to application of boron in boron-deficient calcareous soils. Communication in Soil Science, Plant and Analysis, 38: 1-19. Dell, B., Huang, L. 1997. Physiological response of plants to low boron. Plant and Soil, 193: 103-120. Donald, D.H., Gwathmey, C.O., Carl, E.S. 1998. Foliar feeding of cotton: Evaluating potassium sources, potassium solution buffering and boron. Agronomy Journal, 90: 740-746. Gupta, U.C. (ed.). 1993. Source of boron. In: Boron and its Role in Crop Production. Agriculture and Agri-Food Canada. Charlottetown Saskatchewan, Canada.

121 Gupta, U.C., Jame, Y.W., Campbell, C.A., Leyshon, A.J., Nicholaichuk, W. 1985. Boron toxicity and deficiency: a review. Canadian Journal of Soil Science, 65: 381-409. Hussain, M.Z., Rehman, N., Khan, M.A., Roohullah, Ahmed, S.R. 2006. Micronutrients status of Bannu basen soils. Sarhad Journal of Agriculture, 22: 283-285. Lopez-Lefebre, L.R., Rivero, R.M., Garcia, P.C., Sanchez, E., Ruiz, J.M., Romero, L. 2002. Boron effect on mineral nutrition of tobacco. Journal of Plant Nutrition, 25: 509-522. Mazher, A.M., Zaghloul, S.M., Yassen, A.A. 2006. Impact of boron fertilizer on growth and chemical constituents of Taxodium distichum grown under water regime. World Journal of Agricultural Sciences, 2: 412-420. NNS. 2008. National Nutrition Survey, 2007-2008. Planning Commission, Government of Pakistan, Islamabad, Pakistan. Raun, W.R., Johnson, G.V. 1999. Improving nitrogen use efficiency for cereal production. Agronomy Journal, 91: 357-363. Ruiz, J.M., Baghour, M., Bretones, G., Belakbir, A., Romero, L. 1998. Nitrogen metabolism in tobacco plants (Nicotiana tabacum L.): Role of boron as a possible regulatory factor. International Journal of Plant Sciences, 159: 121-126. Schon, M., Blevins, D. 1990. Foliar boron applications increase the final number of branches and pods on branches of field-grown soybeans. Plant Physiology, 92: 602-605. Shelp, B.J. 1988. Boron mobility and nutrition in broccoli (Brassica oleracea var. italica). Annals of Botany, 61: 83-91. Sherrell, C.G. 1983. Boron deficiency and response in white and red clovers and lucerne. New Zealand Journal of Agricultural Research, 26: 197-203. Shorrocks, V.M. 1997. The occurrence and correction of boron deficiency. Plant and Soil, 193: 121-148. Steel, G.R., Torrie, H.J., Dickey, A.D. 1997. Principals and Procedures of Statistics. A Biometrical Approach. pp. 400-428, 3rd edition, McGraw Hifi Book Co. Inc., New York, USA.

Pak. j. sci. ind. res. Ser. B: biol. sci. 2012 55 (3) 122-128

Diversity Analysis of Marigolds – Tagetes (Asteraceae) Chithra Santhakumari Amma and Rajalakshmi Radhakrishnan* Department of Botany, University of Kerala, Kariavattom, Thiruvananthapuram- 695581 Kerala , India (received December 2, 2011; revised September 6, 2012; accepted September 13, 2012)

Abstract. Characterization of 20 Tagetes cultivars was achieved using morphological and anatomical markers. Tagetes erecta ‘Maurel orange’ (E4) was the most vigorous compared with other genotypes and produced large flowers with high quality and quantity. An anatomical study of leaf secretory cavities differentiated between the cultivars and revealed that foliar secretory cavities were larger and more abundant in T. erecta cultivars than in T. patula cultivars. The size of oil glands also differed in each cultivar. From this study it is concluded that T. erecta cultivars especially T. erecta ‘Maurel orange’ appeared as more promising one to cultivate and more appropriate to be selected as the parental genotype in the hybridization processes for obtaining new varieties and cultivars. It is concluded that different marker systems (morphological, and anatomical) are appropriate to differentiate between the cultivars of Tagetes. Keywords: marigold, Tagetes erecta, Tagetes patula, morphological traits, cultivars, diversity

Introduction

Plant taxonomy has been mainly based upon morphological, cytological, and molecular biological analysis, etc. Morphological characters, both qualitative and quantitative, have long been used to identify species, varieties and cultivars to evaluate relationships, and discriminate between varieties and cultivars. Morphological traits continue to be the first step in the studies of genetic relationships in most breeding programmes (Van Beuningen and Busch, 1997; Cox and Murphy, 1990). In Tagetes breeding programmes, the major emphasis has been on the collection and conservation of genetic pools. There are numerous cultivars that cover a wide spectrum of growth habits, floral traits, environmental responses and varying pest and disease susceptibilities. So a wide range of characters have to be considered to select a superior germplasm that serves as the essential foundation for the breeding of new improved varieties as well as cultivars. Earlier studies on Tagetes cultivars using morphological traits such as plant habit, leaf traits, flower size, flower colour and pigmentation are rare. Anatomical characters for the identification of oil glands are more important to characterize the presence of essential oil in leaves. These traits are found to be of great importance to distinguish genetic variability, and can lead to a better classification of Tagetes cultivars. Thus, the objectives of the present study were to analyze the diversity of 20 cultivars belonging to two species of Tagetes (T.erecta and T. patula) and to classify them into particular groups

Marigolds belong to the family Asteraceae (Compositae), genus Tagetes. Their natural range extends from the southwestern United States into Argentina, with the greatest diversity being in south-central México (Trostle, 1968). The genus Tagetes (Asteraceae) contains 56 species, of which only few species are currently cultivated as horticultural crops. Some companies, release new cultivars every year. Examples are, ‘Marvel’ line, ‘Taishan’line of T. erecta and ‘Bonanza’ line, ‘Boy’ line of T.patula which have been widely used in the world (Zhang et al., 2011). Most of the cultivars were produced in the traditional hybridization breeding way (Tian et al., 2007; Wang, 2003, 2009). Besides, some work has also been done on the breeding of transgenic marigold (Charles et al., 2001). Nowadays, the cultivars widely planted throughout the world belong to three species: T. erecta, T.patula and T. tenuifolia. T. erecta flowers are ideal materials for extracting lutein. Therefore, it is very important to study Tagetes plant dut to its great economic value. Many cultivars have been available however, little is known about the differences between them, possibly because many have been introduced in different places with different names as well as different documentation of the original identities. *Author for correspondence; E-mail: rajalakshmi.murali @gmail.com

122

Diversity Analysis of Marigolds based on morphological and anatomical variation. It can be used for cultivar identification and further use in crop improvement through breeding programmes.

Materials and Methods Plant material. Twelve cultivars of T. erecta and eight cultivars of T. patula were procured from nurseries for this study. 20 genotypes were grown at the Botany Department Garden, Kariavattom, Thiruvananthapuram (Kerala). All plants were replicated by stem cuttings and these vegetative propagated plants (clones) were used for further study. Crop management was done according to recommended agronomic practices. Propagation. Clones were produced through stem cuttings. Plants were grown quickly and produce flowers. For seed germination seeds were sown in pots (germination percentage was only 50%).Out of twenty cultivars only twelve were germinated and rest of it was not being germinated at a stipulated period. The seeds were germinated in 3-5 days. Morphological study. Twenty-cultivars belonging to two species of Tagetes were selected for this study. To avoid phenological differences between individuals, plants that had started to bloom were chosen for the study of vegetative traits and flowers were collected at the anthesis stage. For the study of leaf traits, the third leaf from the apex was chosen. Thirty morphological characters; 20 qualitative and 10 quantitative characters (Table 1), were considerd. Five plants were measured for each trait. The terminology of Hickey and King (2000) was adopted to describe the qualitative characters. Morphological and floral traits were studied with the help of hand lens. Photographs were taken with a digital camera (DP11, Olympus, Tokyo, Japan). A datasheet was designed, and information was recorded for 10 quantitative and 20 qualitative characters (Table 2-3). To evaluate significant differences in quantitative traits, one-way ANOVA was performed. A correlation based PCA was performed to select taxonomically significant qualitative and quantitative characters and to detect outliers. The relative taxonomic distance between the cultivars was calculated by cluster analysis. Pair-wise relationships were estimated by the Euclidean distance coefficient. The distance matrix was represented as a phenogram by the UPGMA clustering method (Sneath and Sokal, 1973). All statistical tests except ANOVA were performed with the MultiVariate Statistical Package

123 Table 1. Morphological characters of Tagetes taken for the study Parameters Qualitative characters Stem surface Petiole Leaf position Leaf Shape Leaf apex Leaf base Leaf margin Leaf colour Lamina symmetry Head type Inflorescence type Inflorescenceposition Involucre type Pappus type Style colour Stigma colour Stigma type Quantitative traits Leaf length Leaf breadth Leaf area Leaf perimeter Peduncle length Internode length Involucre length Pappus length Ovary length Style length

Observation 1. Ribbed; 2. Ribbed and Hairy; 3. Stem Reddish and Ribbed 1.Short; 2. Long 1. Opposite; 2. Alternate 1.Lanceolate; 2.Ovate; 3.Acute 1.Acute; 2. Ovate 1.Acute; 2. Ovate 1.Serrate ; 2. Non serrate 1. Green; 2. Not green 1. Symmetry; 2. Non symmetry 1. Heterozygous; 2. Homozygous 1. Corymbose; 2. Not corymbose 1. Solitary; 2. Axillary 1. Uni serrate; 2. Multi serrate 1. Awns; 2. Capillary; 3. Bristles 1.Yellow; 2. Orange; 3. Pale yellow 4. Pale orange 1.Yellow; 2. Orange; 3. Pale yellow 4. Pale orange 1. Bifid; 2.Not Bifid cm cm cm2 cm cm cm mm mm mm mm

(MVSP) version 3.1 (Kovach Computing Services, Wales, UK). ANOVA was carried out with SPSS 7.5 (SPSS, 1999). Anatomical study. For the study of some anatomical traits, morphologically distinct accessions were selected based on the UPGMA cluster analysis, PCA and oneway ANOVA. They were T. erecta ‘Indian orange’(E1), T. erecta ‘Maurel orange’ (E4), T. erecta ‘Antigua white (E6), T. erecta ‘Discovery yellow’ (E7), T. erecta ‘Antigua Orange’ (E9), T. erecta ‘ Safari Tangerine’ (E11), T. patula ‘Double Eagle’ (P1), T. patula ‘Double Orange’ (P5), T. patula ‘Inca Cream’(P6) and T. patula ‘Safari Yellow’ (P8). For the leaf anatomy analysis, fully developed leaves were collected and fixed in ethanol. Transverse sections of the midrib, as well as

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Table 2. Variations in quantitative characters of Tagetes Morphological Characters Name

Cultivar Leaf code length (cm)

T. erecta‘Indian Orange’

E1

T. erecta‘Atlantis Orange’

E2

T. erecta‘Antigua Yellow’

Leaf breadth (cm)

Leaf area (cm2)

Leaf perimeter (cm)

Peduncle length (cm)

Intermodal Involucres Pappus length length length (cm) (cm) (mm)

6.2 ±0.08 1.6±-0.04

7.3±-0.04

9.2±-0.1

6.3±-0.06 1.3±-0.04 3.2±-0.07

2.2±-0.1

6.3±0.06

1.6±-0.04

7.2±-0.04

9.3±-0

6.1±-0.07 1.2±-0.04 3.1±-0.04

E3

6.5±0.07

1.5±-0.04

7.3±-0.04

9.4±-0.9

6.2±-0.08 1.4±-0.04 3.1±-0.04

2.6±-0.09 1.2±-0.04

2.2±-0.04

T. erecta‘Maurel Orange’

E4

6.4±0.08

1.6±-0.04

7.2±-0.85

8.2±-0.1

5.5±-0.15 1.2±-0.04 3.1±-0.06

2.3±-0.06 1.1±-0.07

2.1±-0.06

T. erecta‘Maurel Yellow’

E5

6.4±0.05

1.5±-0.06

7.1±-0.08

9.2±-0.08 6.3±-0.08 1.4±-0.08 3.1±-0.08

2.6±-0.04 1.2±-0.04

2.3±-0.10

T. erecta‘Antigua White’

E6

6.3±0.04

1.4±-0.04

7.2±-0.6

9.1±-0.6

6.4±-0.1

1.3±-0.04 3.1±-0.08

2.1±-0.06 1.4±-0.10

2.1±-0.07

T. erecta‘Discovery Yellow’ E7

6.5±0.04

1.6±-0.06

7.3±-0.6

9.1±-0.7

6.3±-0.06 1.2±-0.06 3.1±-0.06

2.6±-0.04 1.3±-0.04

2.2±-0.09

T. erecta‘Sweet White’

E8

6.0±0.06

1.5±-0.04

7.3±-0.

9.3±-0.06 6.3±-0.08 1.2±-0.09 3.2±-0.06

2.6±-0.06 1.1±-0.07

2.2±-0.09

T. erecta‘Antigua Orange’

E9

6.6±0.04

1.6±-0.06

7.2±-0.6

9.3±-0.04 6.3±-0.06 1.4±-0.10 3.2±-0.06

2.4±-0.04 1.2±-0.02

2.1±-0.04

T. erecta‘Inca Yellow’

E10

6.4±0.06

1.3±-0.04

7.4±-0.08

9.4±-0.08 6.65±-0.1 1.5±-0.08 3.1±-0.06

2.4±-0.04 1.4±-0.06

2.1±-0.06

T. erecta‘ Safari Tangerine’ E11

6.4±0.07

1.2±-0.07

7.2±-0.7

9.2±-0.06 6.5±-0.1

2.7±-0.01 1.3±-0.04

2.1±-0.04

T. erecta‘ Sweet Cream’

E12

6.6±0.06

1.4±-0.04

7.4±-0.04

9.4±-0.1

6.3±-0.07 1.2±-0.10 3.3±-0.08

2.5±-0.04 1.3±-0.06

2.2±-0.0

T. patula ‘Double Eagle’

P1

5.3±0.07

1.2±-0.08

6.9±-0.07

8.3±-0.08 5.0±-0.14 1.2±-0.00 2.1±-0.04

3.0±-0.08 2.1±-0.06

2.1±-0.06

T. patula‘Inca Orange’

P2

5.4±0.08

1.2±-0.08

7.1±-0.06

8.3±-0.07 5.3±-0.08 1.2±-0.07 2.3±-0.04

3.1±-0.06 2.2±-0.09

2.2±-0.09

T. patula ‘Orange Eagle’

P3

5.3±0.08

1.3±-0.06

7.1±-0.06

8.2±-0.10 5.4±-0.09 1.2±-0.08 2.1±-0.06

3.2±-0.06 1.1±-0.04

2.1±-0.04

1.4±-0.06 3.4±-0.1

Ovary length (mm)

Style length (cm)

1.4±-0.04

2.1±-0.06

2.2±-0.05 1.2±-0.08

2.1±-0.04

T. patula ‘Safari’

P4

5.4±0.08

1.4±-0.04

7.2±-0.02

8.2±-0.09 5.6±-0.06 1.2±-0.04 3.2±-0.00

2.1±-0.06 1.3±-0.06

1.3±-0.06

T. patula ‘Double Orange’

P5

5.2±0.04

1.1±-0.06

6.9±-0.04

8.4±-0.11

2.1±-0.06 1.2±-0.07

1.2±-7.07

T. patula ‘Inca Cream’

P6

5.7±0.05

1.3±-0.06

6.9±-0.04

8.3±-0.06 5.3±-0.00 1.2±-0.08 3.0±-0.06

2.4±-0.02 1.1±-0.0

1.1±-0.08

T. patula ‘Mesa Gold’

P7

5.3±0.09

1.0±-0.06

7.0±-0.08

8.5±-0.07 5.3±-0.09 1.2±-0.08 3.0±-7.5

2.2±-0.08 1.1±-0.06

1.1±-6.29

T. patula‘Safari Yellow’

P8

5.2±-0.08 1.3±-0.02

6.7±-0.06

8.4±-0.08 5.6±-0.04 1.2±-0.08 3.1±-0.08

2.1±-0.06 1.1±-0.06

1.1±-6.29

5.6±-0.06 1.3±-0.04 3.2±-0.01

Table 3. Qualitative characters of Tagetes Cultivars Characters

E1

E2

E3

E4

E5

E6

E7

E8

E9

E10 E11 E12 P1

P2

P3

P4

P5

P6

P7

P8

Habit Stem surface Branching nature Petiole Leaf position Leaf shape Leaf apex Leaf base Leaf margin Leaf colour Lamina symmetry Flower colour Head type Inflorescence type Inflorescence position Involucre type Pappus type Style colour Stigma colour Stigma type

H RH SB L OP OV AC AC SE G SY O HO CO AX UN CB y Y BF

H R PB L OP LA AC AC SE G SY LO HO CO AX UN CB PY Y BF

H R NB L AL LA AC AC SE G SY PY HO CO SO UN CB Y Y BF

H RH PB L OP LA AC OV SE G SY DO HO CO AX UN CB PO PO BF

H RH NB L AL OV AC AC SE G SY FY HO CO AX UN CB Y Y BF

H RH NB L AL AC OV OV SE G SY WH HO CO SO UN CB O O BF

H RH SB L OP AC OV AC SE G SY DY HO CO SO UN CB WH WH BF

H RH SB L OP AC OV OV SE G SY CW HO CO SO UN CB O O BF

H R SB L AL AC OV OV SE G SY PO HO CO AX UN CB WH WH BF

H R PB L OP AC OV OV SE G SY LY HO CO AX UN CB O O BF

H RR B M OP OV AC AC SE G SY YO HE CO SO MU AW O O BF

H RH PB S AL OV AC OV SE G SY DO HE CO SO MU AW PY PY BF

H RR B S AL OV AC AC SE G SY DY HE CO SO MU AW O O BF

H R PB S AL OV AC AC SE G SY DR HE CO AX MU AW PO PO BF

H RR PB S AL OV AC AC SE G SY YC HE CO CO MU AW PO PO BF

H RR PB S OP AC OV OV SE G SY GY HE CO CO MU AW Y Y BF

H RR PB S OP OV AC AC SE G SY DY HE CO CO MU AW O O BF

H RR PB L AL AC OV AC SE G SY YO HO CO SO UN CB Y Y BF

H RH PB L OP AC OV AC SE G SY C HO CO SO UN CB O YO BF

H R B S OP LA AC OV SE G SY YR HE CO SO MU AW Y Y BF

R = Ribbed; RH = Ribbed and Hairy; RR = Stem Reddish and Ribbed; SB = Sparingly branched; PB = Profusely branched; NB = Not branched; S = Short; L = Long; OP = Opposite; AL = Alternate; LA = Lanceolate; OV = Ovate; AC = Acute; S = Serate; G = Green; S = Symmetry; HE = Heterozygous; HO = Homozygous; C = Corymbose; SO = Solitary; AX = Axillary; UN = Uni serrate; MU = Multi serrate; AW = Awns; CB = Capillary bristles; Y = Yellow; O = Orange; PY = Pale yellow; PO = Pale orange; BF = Bifid.

Diversity Analysis of Marigolds transverse sections of the leaf-blade in the intercostal region and in the margin were obtained. Hand-sections were performed using a razor blade, cleared with sodium hypochlorite, washed with water, stained with astrablausafranin (Bukatsch, 1972) and mounted in 50% glycerol. Anatomical features were studied by using a stereo zoom microscope (SZ61, Olympus, Tokyo, Japan). Photographs were taken with a digital camera (DP11, Olympus, Tokyo, Japan) attached to the microscope.

Results and Discussion Morphological study. Morphological characters. The plants were herbs and the shoot length of T.erecta ranged between 12 to 16 cm, while the shoot length of T. patula ranged between 8 to 10 cm. Stem colour was reddish or green and stem surface was ribbed or ribbed and hairy. Some genotypes were unbranched as E3, E5 and E6 while the other genotypes were sparingly or profusely branched. Internodal length varied from 1.2 to 1.5 cm in T. erecta and 1.2 to 1.3 cm in T. patula. The leaves were green, compound and symmetrical, lanceolate, acute or ovate with serrate margin. Leaves were opposite or alternate. Leaf length ranged from 6.2 to 6.6 cm in T. erecta and 5.2 to 5.7 cm in T. patula. Leaf width ranged between 1.2 to 1.6 cm in T. erecta and 1.0 to 1.4 cm in T. patula. Leaf area and perimeter in T. erecta ranged between 7.1 to 7.4 cm2 and 8.2 to 9.4 cm respectively. In T. patula it ranged between 6.7 to 7.2 cm2 and 8.2 to 8.5 cm respectively. The inflorescence consists of numerous ray and disc florets in head or capitulum. Head type was homogamous in T.erecta but herterogamous in T. patula. Variations could be noted in the flower colour. It was orange in E1, E2, E4 and E9. Various yellowish shades were seen in E3, E5, E7, E10 and E11. White, creamy white and cream colours were found in E6, E8 and E12 respectively. Some cultivars of T.patula showed distinct colours in outside and inside of the head inflorescence. Reddish yellow outside and yellowish inside in P1, orange outside and yellow inside in P3 and whitish cream outside and yellow inside in P6. Dark red coloured ray florets were found in P5. Involucre type in T.erecta was uniserrate but in T. patula it was multi serrate. Capillary bristles were found in T. erecta but awns in T. patula. Involucre length of T. erecta ranged between 3.1 to 3.4 cm where as in T. patula it ranged between 2.1 to 3.2 cm. Pappus length of flowers ranged between 2.1 to 2.7 cm in T. erecta and 2.1 to 3.2 cm in T. patula. Ovary and style length of T. patula were found to be

125 more variable compared to T. erecta. In T. patula ovary length ranged between 1.1 to 2.2 cm where as in T. erecta it was 1.1 to 1.4 cm. Style length ranged between 1.1 to 2.2 cm in T. patula where as in T. erecta it was 2.1 to 2.3 cm. Stigma was bifid in both species. Morphometric analysis. ANOVA. All the quantitative characters were found to be significant. PCA. Qualitative traits. In the PCA of qualitative data of T.erecta, 56.8% of the phenetic variance was accounted for by the first principal axis, followed by 17.3% for the second, 8.2% for the third, 5.5% for the fourth and 5.3% for the fifth principal component (Table 4). Most of the selected qualitative traits were found principally influential in the PCA. Branching nature, leaf morphology and flower colour were found influential in the most variable first principal component. In the PCA of qualitative data of T.patula, 38.6% of the phenetic variance was accounted for by the first principal axis, followed by 25.5% for the second, 16.2% for the third, 8.6% for the fourth and 6.9% for the fifth principal component (Table 4). Most of the selected qualitative traits were found principally influential in the PCA. Leaf position, leaf morphology and flower colour were found influential in the most variable first principal component. Quantitative traits. In the PCA of quantitative data of T.erecta, 43.3% of the phenetic variance was accounted for by the first principal axis, followed by 23.2% for the second, 15.8% for the third and 10.3% for the fourth principal component (Table 5). The first principal component accounted for 62.7% of phenotypic variance and the second component for 21.1% (Table 5) in T.patula. All the quantitative traits except leaf characters were found to have significant loadings in PCA. Cluster analysis. The UPGMA phenogram based on the morphological characters shows two main principal clusters (Fig. 1) separating the two species. In the T.patula cluster, the cultivars P3 and P4 are separated out with others, and again P2 and P6 are separated out. Cultivar P7 is more distinct from P2, P6, P3 and P4 but more close to P8, P1 and P5. In the T. erecta cluster cultivar E4 is more distant from others. E8 and E7 as well as E10 and E11 are more close to each other. Anatomical study. Anatomical studies revealed that foliar secretory cavities were more abundant in T. erecta cultivars than in T. patula cultivars. The secretory structures were found in the lamina between the palisade

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Rajalakshmi Radhakrishnan and Chittra Santhakumari Amma UPGMA

0.15

0.125

0.1

P7 P8 P1 P5 P2 P6 P3 P4 E4 E11 E10 E6 E5 E3 E2 E1 E9 E12 E8 E7

0.075

0.05

0.025

0

Euclidean - Data log (10) transformed

E1 E2 E3 E4 E5 E6 E7 E8 E9 E10 E11 E12

T. erecta ‘Indian Orange’ T. erecta ‘Atlantis Orange’ T. erecta ‘Antigua Yellow’ T. erecta ‘Maurel Orange’ T. erecta ‘Maurel Yellow’ T. erecta ‘Antigua White’ T. erecta ‘Discovery Yellow’ T. erecta ‘Sweet White’ T. erecta ‘Antigua Orange’ T. erecta ‘Inca Yellow’ T. erecta ‘ Safari Tangerine’ T.erecta ‘ Sweet Cream’

P1 P2 P3 P4 P5 P6 P7 P8

T. patula ‘Double Eagle’ T. patula ‘Inca Orange’ T. patula ‘Orange Eagle’ T. patula ‘Safari’ T. patula ‘Double Orange’ T. patula ‘Inca Cream’ T. patula ‘Mesa Gold’ T. patula ‘Safari Yellow’

Fig. 1. UPGMA phenogram Tagetes based on morphological characters. Table 4. PCA variable loading of morphology, (qualitative) in Tagetes cultivars Anatomical features

Stem surface Branching nature Petiole Leaf position Leaf Shape Leaf apex Leaf base Leaf margin Leaf colour Lamina symmetry Flower colour Head Type Inflorescence type Inflorescene position Involucre type Pappus type Style colour Stigma colour Stigma type Eigenvalues Percentage Cum. Percentage

Tagetes erecta

Tagetes patula

Axis 1

Axis 2

Axis 3

Axis 4

Axis 5

Axis 1

Axis 2

Axis 3

Axis 4

Axis 5

-0.058 -0.148 0.000 -0.070 0.307 0.267 0.196 0.000 0.000 0.000 0.383 0.000 0.000 0.080 0.000 0.000 0.584 0.521 0.000 2.639 56.784 56.784

-0.470 -0.370 0.000 -0.382 -0.259 -0.200 0.046 0.000 0.000 0.000 -0.549 0.000 0.000 0.058 0.000 0.000 0.246 0.148 0.000 0.804 17.305 74.089

-0.458 0.703 0.000 0.274 -0.084 -0.128 0.184 0.000 0.000 0.000 -0.058 0.000 0.000 0.374 0.000 0.000 0.101 0.104 0.000 0.383 8.244 82.333

-0.112 0.341 0.000 -0.265 -0.235 -0.170 0.166 0.000 0.000 0.000 0.225 0.000 0.000 -0.782 0.000 0.000 0.164 -0.016 0.000 0.256 5.504 87.837

0.560 0.138 0.000 0.059 0.206 -0.200 0.521 0.000 0.000 0.000 -0.489 0.000 0.000 -0.033 0.000 0.000 0.261 -0.034 0.000 0.248 5.342 93.179

0.077 0.077 -0.019 0.378 0.121 -0.130 0.247 0.000 0.000 0.000 0.223 0.000 0.000 0.259 0.000 0.000 0.484 0.634 0.000 1.403 38.640 38.640

0.397 0.397 -0.066 -0.048 0.346 0.195 0.029 0.000 0.000 0.000 0.618 0.000 0.000 -0.063 0.000 0.000 -0.365 -0.021 0.000 0.927 25.540 64.180

-0.466 -0.466 -0.175 -0.041 -0.064 0.021 0.246 0.000 0.000 0.000 0.392 0.000 0.000 0.373 0.000 0.000 -0.411 0.076 0.000 0.587 16.168 80.348

0.209 0.209 0.263 -0.088 -0.275 -0.393 0.444 0.000 0.000 0.000 -0.275 0.000 0.000 -0.002 0.000 0.000 -0.500 0.289 0.000 0.313 8.627 88.975

-0.115 -0.115 0.497 -0.465 0.288 0.068 0.507 0.000 0.000 0.000 0.130 0.000 0.000 -0.142 0.000 0.000 0.325 -0.154 0.000 0.251 6.913 95.888

Diversity Analysis of Marigolds

127

Table 5. PCA variable loading of morphology, (quantitative) in Tagetes cultivars Anatomical features Leaf length Leaf breadth Leaf area Leaf perimeter Petiole length Internode length Involucre length Pappus length Ovary length Style length Eigenvalues Percentage Cum. Percentage

Tagetes erecta

Tagetes patula

Axis 1

Axis 2

Axis 3

Axis 4

Axis 1

Axis 2

-0.006 0.649 -0.002 -0.229 -0.240 -0.515 -0.030 -0.440 -0.078 0.057 0.012 43.266 43.266

0.050 0.590 0.035 0.194 0.219 0.017 -0.101 0.692 -0.153 0.222 0.007 23.210 66.476

-0.292 0.403 0.014 0.083 0.185 0.679 -0.239 -0.373 0.193 -0.128 0.005 15.845 82.321

-0.154 -0.052 0.101 0.330 0.561 -0.401 0.070 -0.154 0.543 0.235 0.003 10.372 92.693

-0.084 0.004 0.040 0.001 0.085 -0.344 0.465 -0.396 0.646 -0.275 0.017 62.740 62.740

0.169 0.648 0.183 -0.017 -0.050 -0.139 0.577 0.138 -0.284 0.253 0.006 21.126 83.867

and spongy parenchymatic tissue and in the cortical region of the petiole. The size of oil glands differed among the cultivars. In the cultivars of T. erecta the oil glands were larger in size compared with the oil glands in T. patula cultivars. In the present study, an analysis of morphological characters of Tagetes species revealed that each cultivar is distinct. The two species of Tagetes (T. erecta and T. patula) used for this study were morphologically dissimilar. The shoot length of T. erecta ranged between 12 to 16 cm, while the shoot length of T. patula ranged between 8 to 10 cm suggests that T. patula plants were smaller than T. erecta. Branching nature was also different. Most of the cultivars from T. patula were shown profusely branched nature. Variations could be noted in the flower colour. It was orange in T. erecta ‘Indian orange’(E1), T. erecta ‘Atlantis orange’ (E2), T.erecta ‘Maurel orange’ (E4), T. erecta ‘Antigua Orange’ (E9) and T. erecta ‘Safari Tangerine’ (E11). Various yellowish shades were seen in T.erecta ‘Antigua yellow’ (E3), T. erecta ‘Maurel yellow (E5), T. erecta‘ Discovery yellow’ (E7) and T. erecta ‘ Inca Yellow’ (E10).White, creamy white and cream colours were found in T. erecta ‘Antigua white (E6), T. erecta ‘Sweet white’ (E8) and T. erecta‘ Sweet Cream’ (E12) respectively. The most attractive flowers were observed in T. patula and head was heterogamous. The biggest flowerings form at T. erecta (Tagetes erecta ‘Maurel Orange’[E4]) and smallest ones at T.patula (T.patula ‘Inca Orange’ [P2]).

Cluster analysis substantiated the existence of diversity among the 20 cultivars for the morphological traits studied. The clustering pattern showed that two principal clusters for two species, T. erecta ‘Maurel orange’ (E4) was distant from other cultivars of T. erecta. The dendrogram constructed on the basis of the data generated from the morphological traits of T. erecta also showed E4 was distinct from others. The colour of the flower was more attractive in this cultivar and other notable characters were the ribbed and hairy stem surface, long petiole, opposite leaves with serrate margin, leaf shape lanceolate and lamina symmetrical. Peduncle was found to be short (5.5 cm) compared to other cultivars. Biggest flowers with dark orange colour confirm its high pigment content rather than attractiveness. Anatomical characterization of leaf showed the presence of large oil glands in this cultivar. Both these characters make up this cultivar more economical. Anatomical studies revealed that foliar secretory cavities were more in T. erecta cultivars than in T. patula cultivars. The size of oil glands differ in each cultivar. In the cultivars of T. erecta, the oil glands were larger in size compared with the oil glands in T. patula cultivars. From this present study it is clear that T. erecta cultivars are more appropriate to select as the parental genotype in the hybridization processes for obtaining different varieties and cultivars. Since many reports show that T. erecta are good source of yellow pigments and essential oil content useful for various activities (Verghese,1998). Ethnobotanical studies on T. erecta were also reported by many workers (Balick

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Rajalakshmi Radhakrishnan and Chittra Santhakumari Amma

and Cox, 1996; Neher, 1968;). Vasudevan et al. (1997) has ranked T. erecta as a multipurpose herb. A marigold flower contains abundant amount of a valuable antioxidant compound called lutein (Lavececchia et al.,2004). The extract with only the purified form with a lutein content of known concen-tration and a pure crystalline lutein isolated from marigold flower especially from T. erecta is allowed for food use. Dark coloured flowers contain about 200 times more lutein than the light coloured flowers. The concentration of lutein varies in different shades of marigold flowers, viz.; greenish yellow to bright yellow and orange brown (Gregory et al., 1986). The dark orange colour flowers are observed in T. erecta cultivars. From the present study it was evident that T. erecta ‘Maurel orange’ (E4) have large and dark orange coloured flowers. Because of the wide range of activity marigolds are cultivated by farmers, but in India mostly for its yellow flowers. After harvesting they destroy the plants without utilizing its full benefits. Each and every part of the plant is very much useful. Simple distillation of the plant parts achieve good amount of essential oil which can be used for various purposes. Still the identification of superior cultivar is a difficult task. Various characters are spread in different cultivars and breeders can not find these characters in a single plant. Different genes which are distributed in different cultivars if pooled together in single plant will definitely create a superior plant which can be achieved through breeding. This study of preliminary screening of these cultivars by morphological and anatomical traits revealed that T. erecta cultivars are more appropriate to be selected as the parental genotype in the hybridization processes. Among this T.erecta ‘Maurel orange’ (E4) is superior in floral traits as well as large oil glands. It is concluded from these results that different marker systems are appropriate to differentiate between the cultivars of Tagetes. Also, these marker systems could be complementary to each other and should be followed by molecular characterization using PCR-based markers to establish an integrated data about these cultivars. The maximum variability was present in flower colour. The cultivars with dark orange colour flowers can be used in further hybridization programme because of high carotenoid content. Selection of better cultivar can be made for species or varietal improvement on the basis of percent similarity with other species. Two more prominent characters, i.e., large oil glands and dark orange flowers can be chosen for the purpose.

References Balick, J.M., Cox, P.A. 1996. Plants, people and culture: In: The Science of Ethnobotany. 228 pp. Scientific American Library, New York, USA. Bukatsch, F. 1972. Bermerkungen zur Doppelfärbung Astrablau-Safranin. Mikrokosmos, 61: 255. Charles, P.M., Li, T., Katherine, W.O. 2001. Analysis of carotenoid biosynthetic gene expression during marigold petal development. Plant Molecular Biology, 45: 281-293. Cox, T.S., Murphy, J.P. 1990. The effect of parental divergence on F2 heterosis in winter wheat crosses. Theoretical and Applied Genetics, 79: 241-250. Gregory, G.K., Chen, T.S., Philip, T. 1986. Quantitative analysis of lutein esters in marigold flowers by high performance liquid chromatography. Journal of Food Science, 51: 1093-1094. Hickey, M., King, C. 2000. The Cambridge Illustrated Glossary of Botanical Terms. P. R. Shewry (ed.), vol. 35, 208 pp., Cambridge University Press, Cambridge, UK. Lavececchia, C. 2004. Mediterranean diet and cancer. Public Health and Nutrition, 7: 965-968. Neher, R.T.1968. The ethnobotany of Tagetes. Economic Botany, 22: 317-325. Sneath, P.H.A., Sokal, R.R. 1973. Numerical taxonomy, In: The Principles and Practice of Numerical Classification, pp. 573-576, W.H. Freeman and Company, San Francisco, USA. SPSS. 1999. Manuals SPSS Inc., Chicago, V-10.0. Tian, H.Y., Wang, P., Shen, X.Q. 2007. Genetic analysis and botanical character in male sterile W205AB line of marigold. Northern Horticulture, 2: 105-107. Trostle, R. 1968. The ethnobotany of Tagets Economic Botany, 22: 317-325. Van Beuningen, L.T., Bush, R.H. 1997. Genetic diversity among North American spring wheat cultivars: I. Analysis of the coefficient of parentage matrix. Crop Science, 37: 570-579. Vasudevan, P., Suman, K., Satyavathi, S. 1997. Tagetes: A multipurpose plant. Bioresource Technology, 62: 29-35. Verghese, J. 1998. Focus on xanthophylls from Tagetes erecta L., the giant natural color complex-II. Indian Spices, 34: 13-16. Wang, P. 2009. Selection and application of a new pigment Tagetes erecta 'Sesu 1'. Liaoning Agricultural Science, 2: 74-75. Wang, G.Y. 2003. Technology on production of hybrid marigold seed in protected field. China Seed Industry, 10: 59-60. Zhang, P., Zeng, L., Su, Y., Gong, X., Wang, X. 2011.Karyotype studies on Tagetes erecta L. and Tagetes patula L. African Journal of Biotechnology, 10: 16138-16144.

Pak. j. sci. ind. res. Ser. B: biol. sci. 2012 55 (3) 129-137

A Study of Poisonous Plants of Islamabad Area, Pakistan Saleem Ahmad

Pakistan Museum of Natural History, Garden Avenue, Islamabad, Pakistan (received June 15, 2010; revised October 1, 2012; accepted October 8, 2012)

Abstract. Poisonous plants growing wild and cultivated in Islamabad area were studied. A total of 45 taxa belonging to 30 families are reported here from the study area. These include 6 trees, 12 shrubs and 27 herbs. Twenty species are found wild in the area, another 20 species are cultivated, while 5 species are found both cultivated and naturally occurring. Besides taxonomic details, poisonous parts, chemical constituents responsible for poisoning, and the human specific physiological effects of poisoning in relation to each plant are presented in a tabular format. The harm caused by these poisonous plants is often not serious, and is primarily restricted to gastrointestinal irritation or mild nervous system effects, that is curable. However, there have been cases of death resulting from the consumption of parts of highly poisonous plants in the area. Keywords: poisonous plants, vegetation taxonomy, environment, Islamabad

Introduction

and the central nervous system (Frohne and Pfander, 1984). Families like Amaryllidaceae, Apocynaceae, Buxaceae, Asteraceae, Euphorbiaceae, Fabacae, Liliaceae, Papaveraceae, Ranunculaceae and Solanaceae are particularly rich in alkaloids.

A strict definition of a poisonous plant is not easily constructed, since the boundaries between harmful and harmless plants are frequently blurred. A poisonous plant is usually defined as one whose consumption in a certain amount gives rise to abnormal or impaired functions in the human physiology (Cooper and Johnson, 1984; Lewis and Elvin-Lewis, 1977). These effects are due to the presence of certain toxic chemicals within a plant, which may be ingested, injected, absorbed or contacted through the skin. Some plants have evolved secondary metabolites through the course of evolution, which are not directly involved in their typical growth functions, but rather they serve a protective role for plant. For instance, presence of some of these substances makes a plant slightly to highly poisonous for animals, thus serving as a deterrent to herbivory.

Glucosides and glycosides form a group of chemicals that upon splitting give rise to a carbohydrate and one or more other products called aglycones. Although many of these chemicals are not toxic, some cause poisoning by releasing toxic chemicals on hydrolysis (e.g. cyanide or hydrocyanic acid). The majority of cyanide poisoning is caused by species belonging to the Rosaceae, Fabaceae and Poaceae. The cyanogenic glucoside amygdalin is found in bitter almonds and commonly causes cyanide poisoning. Cardiac glycosides are known for their anti-arrhythmic effects on the heart as well as their high levels of toxicity, and are found in taxa such as foxglove (Digitalis spp.) and oleander (Nerium oleander L.). Bitter principles are neutral substances present in some Cucurbitaceae, Aloe, Cassia, Artemisia, etc. which have a nauseous, bitter taste and purgative action. Phenols are small molecules containing one or more phenolic group. They are the most widely distributed class of secondary metabolites with several thousand different compounds identified. Most of the phenols are known to have an antioxidant activity (Dimitrios, 2006).

Poisonous plants can be classified on the basis of the chemicals they possess such as alkaloids, glycosides (including saponins), nitrates, bitter principles, oxalates, tannins, phenols, and volatile oils. Alkaloids are the most important group of chemical bases that have an alkali-like reaction, with over 4000 distinct compounds that have been discovered to date. Nicotine, colchicine, morphine, ephedrine, and atropine (among others) are common alkaloids that are most familiar to the general public. These have a bitter taste and show pharmacological activity that often effect the gastrointestinal tract

Oxalates found in the form of oxalic acid and its salts are common in many plants but their abundance causes

E-mail: [email protected]

129

130 toxic effects, especially in members of the Oxalidaceae, Geraniaceae, Araceae and Polygonaceae. Nitrogenous compounds including certain proteins, peptides and amino acids also cause poisoning and are found in certain plants. Toxic proteins or toxalbumins have been found in Fabaceae, Euphorbiaceae, etc. and cause agglutination of the red blood corpuscles. Essential or volatile oils, which impart the characteristic odour in plants have sharp burning taste and can be toxic internally affecting the GIT and CNS. These are abundantly present in Lamiaceae, Asteraceae, Rutaceae, Apiaceae, Myrtaceae, etc. Tannins are non-nitrogenous derivatives having an astringent action, some of which are toxic if taken in large amount. These are common in many plants. Resins form a heterogeneous group of complex and variable chemical substances. Some of these cause poisoning due to their purgative or irritant properties as in certain species of the Cucurbitaceae, Convolvulaceae and Euphorbiaceae (Tampion, 1977; Nasir and Ali, 1970; Dymock et al., 1890-1893). Islamabad is situated on the north eastern part of the Potohar plateau and is located between latitudes 33 °36’ and 33 °49’ N and longitudes 72 °20’ and 73 °24’ E. It covers an area of 382 km 2 of undulating topography and the altitude varies from 503 to 610 m. The mean maximum and minimum temperatures in the summer are 34.2 °C and 24.4 °C; those in winter are 16.7 °C and 3.4 °C, respectively. Rainfall occurs in the monsoon season and winter, with the annual average being 1143 mm. Rawal and Simli Dams are the main water reservoirs, and several small streams are running from north to south in this area. The original vegetation of the site is subtropical scrub forest type (Champion et al., 1965), which has been considerably modified by the introduction of large number of cultivated trees (Ahmad and Khattak, 2001). The flora of the area is an extension of the Mediterranean type (Stewart, 1957). Among all the cities of Pakistan, Islamabad has the richest vegetation, both in terms of the total number of plants as well as the number of species. The green belts provide habitat for not only many cultivated taxa, but also for a large number of wild herbs and grasses which thrive throughout the seasons (Ahmad and Khattak, 2001). While most of these plants have a beneficial effect on the environment of the area, a few harmful species also get the chance to grow and proliferate. Because of their proximity with the population, they are certainly likely to come with human contact and

Saleem Ahmad

may lead to poisoning if consumed out of inquisitiveness or a self-medication effort. Some pertinent references on poisonous plants have been published in the USA and Europe (Connor, 1997; Cooper and Johnson, 1984; Frohne and Pfander, 1984; Tampion, 1977; Kingsbury, 1964). There has also been ample discussion of poisonous plants in several books and encyclopaedias dealing exclusively with medicinal plants (Heywood and Chant, 1982; Grieve, 1979; Stuart, 1979; Lewis, and Elvin-Lewis, 1977; Anderson, 1967). Likewise, regionally focused information on Indian poisonous plants have been provided in several Indian literature sources (Chopra et al., 1984, 1956; Pandey, 1984; Modi, 1945; Dymock, et al., 1890-1893). In contrast, only a few references are available on the specific poisonous plants of Pakistan (Baquar, 1989; Ikram and Hussain, 1978; Baquar and Tasnif, 1967).

Materials and Methods Specimens of plants known to cause poisoning were collected from various areas of Islamabad during 20082009 and prepared following standard herbarium procedures. Specimens were identified with the help of floras and manuals (Polunin and Stainton, 1985; Willis and Airy Shaw, 1985; Stewart, 1972; Nasir and Ali, 1970; Hooker, 1875-1888). Voucher specimens were deposited in the Herbarium of the Pakistan Museum of Natural History (PMNH). Rare specimens were consulted in the Herbarium of Quaid-i-Azam University, Islamabad and National Herbarium Rawalpindi, Pakistan. All taxa were listed in tabular format, and include the following information: scientific, common, and local names, occurence, growth habit and duration, poisonous parts, chemicals constituents responsible for poisoning, and the human specific physiological effects of poisoning in relation to each plant.

Results and Discussion The poisonous plants of Islamabad are summarized in Table 1. It gives knowledge of poisonous plants at a glance and severity of poison of each plant included in this list. Yellow Oleander (Thevetia peruviana) was planted in gardens and roadsides in Islamabad for its attractive flowers. But, its fruit is poisonous, consumption has lead to multiple deaths. Lantana or Panch Phuli (Lantana camara) was planted as an ornamental bush in Islamabad, but has spread very much and formed

Dhak, Flame of the Forest

Ak, Milkweed

Bhang, Charas, Indian Hemp, Marijuana Mirch, Chilies, Cayenne

Sada Bahar, Madagasar Periwinkle

Butea monosperma (L.) Taub. (Papilionaceae)

Calotropis procera (Ait.) Ait.f. (Asclepiadaceae)

Cannabis sativa L. (Cannabaceae)

Capsicum frutescens L. (Solanaceae)

Catharanthus roseus (L.) G. Don (Apocynaceae)

The latex

All parts, especially the seeds

Perennial herb bearing pink flowers

Perennial herb

Leaves

Fruits and seeds

Indole alkaloids vinblastine and vincristine

The pungent principle capsaicin

Effects the heart, acting as a cardiac poison.

Large doses cause difficulty of swallowing, inflammation of the oesophagus and stomach; causing locally skin irritation.

Firstly excitement, visual hallucination, rapid pulse, euphoria, secondly narcosis with dilated pupils.

Internally digestive disorders, nausea and kidney-inflammation. Conjunctivitis and harm to the cornea.

Cont’d ........

Cultivated as an ornamental.

Cultivated for consumption.

Very common wild herb.

In dry places.

Foothills of Margalla. Retching, pain in the abdomen, and occasionally vomiting and giddiness.

A fixed oil in the seeds and glucoside butrin of the flower sap A neutral principle calotropin

Cultivated in gardens and selfgrown.

Constriction of blood vessels latter; dyspnoea, vomiting and diarrhea.

Asclepione, which contains 3 cardioactive glucosides

Often cultivated in homes and gardens.

Diarrhea, menorrhagea, diuresis.

A common wild herb.

Occurence

Harmful effects

Taken internally it produces intense headache and nausea.

Glucosidic saponins

Erect annual herb Flowering shoots, Tetrahydrocannawith small, green leaves, especially binols, found in the flowers of female plant resin

Undershrub with purple flowers

Wild tree in the foothills

Entire plant, especially juice and roots

Kakatundi, Blood weed, Silk weed,

Asclepias curassavica L. (Asclepiadaceae)

Erect shrub with small flowers

Small annual herb Entire plant with blue flowers

Dhabbar, Blue Pimpernell

Anagallis arvensis L. (Primulaceae)

Acrid volatile oil, oxalic acid, sapono-side

Roots and leafjuice

Perennial herb with spine-tipped leaves

Vilayati Kaitalu, Century Plant

Agave americana L (Agavaceae)

Chemical constituents

Parts causing harm

Growth Habit and Duration

Local / Common Name

Scientific name and family

Table 1. Summary of poisonous plants of Pakistan (in alphabetical order by genus).

Poisonous Plants of Islamabad

131

Small annual with root tuber & yellow flowers Erect annual herb bearing blue flowers Erect herb bearing small, green flowers Bushy herbs bearing large white flowers

Perennial herb bearing large, variegated leaves Bushy shrub with small, blue flowers

Tumba, Indrayan, Bitter Apple, Colocynth Surinjan talkh, Meadow Saffron Larkspur

Paleet, Canadian Fleabane Dhatura, Thorn Apple

Dumb Cane

Golden Dewdrop Great Hairy Willow Herb

Citrullus colocynthis Schrad. (Cucurbitaceae)

Colchicum luteum Baker (Colchicaceae)

Consolida ambigua (L.) Ball & Heywood (Ranunculaceae)

Conyza canadensis (L.) Cronq. (Asteraceae)

Datura fastuosa L D. innoxia Miller D. stramonium L. (Solanaceae)

Diffenbachia seguine Schott (Araceae)

Duranta repens L. (Verbenaceae)

Epilobium hirsutum L. (Onagraceae)

Erect herb with small, pink flowers

Prostrate annual herb with yellow flowers

Bushy shrub

Raat ki Rani, Night Jessamine

Cestrum nocturnum L (Solanaceae)

Prostrate herb with round leaves

Brahmi Booti, Indian Pennywort

Centella asiatica (L.) Urban (Apiaceae)

Table 1 Cont’d)

Entire plant

Fruits

Plant juice

Tannin

A saponin

Calcium oxalate crystals

All parts especially Alkaloids juice and seeds hyoscyamine, hyoscine

An essential oil called oil of fleabane

Epileptiform convulsions.

Insomnia, fever and convulsions, sometimes fatal.

Inflamed mouth, swallowing difficulty, swelling of tongue, inability to talk.

Burning stomach pain, lack of coordination, fever, dilated pupils, weak pulse, delirium, hallucinations, respiratory failure.

Dermatitis on external contact; taken internally sore throat, aching of extremities and colic.

Occasional on fallow land. Cont’d ........

Cultivated as hedge plant.

Cultivated as ornamental.

Occasionally found on disturbed or fallow land.

Very common in disturbed or unused places.

Commonly cultivated as ornamental. Vomiting, abdominal pain, blurred vision, agitation, dilation of pupils, respiratory paralysis.

Alkaloids delphinine and ajacine

All parts, particularly the seeds The leaves

Occasionally found in early spring.

Throughout the drier parts of the country.

Commonly cultivated as an ornamental.

In wet places.

Burning of mouth and throat, abdominal pain, violent vomiting, diarrhoea, shock, collapse.

Violent gripping, prostration, sometimes bloody discharge resulting in death.

Nervous and muscular excitement, hallucinations, tachycardia, salivation, dyspnoea, paralysis.

In large doses it is a stupefying narcotic, producing giddiness, vertigo and coma.

Alkaloid colchicine

Resinous material elatrin, phytosterol hentra aconit.

Water-soluble steroidal glycosides

A bitter principle vellarine, resin, indocentello-side, asiaticoside

All parts, particularly the corm and seeds

Pulp of the fruit

All parts

Entire plant

132 Saleem Ahmad

Tamatur, Tomatto

Bakain, Dhrek Persian Lilac

Gul-e-Abbas Four O’ Clock

Nargis, Narcissus

Kaner, Oleander

Khatti booti, Amlika, Yellow Sorrel

Post, Opium poppy

Congress weed

Lycopersicon esculentum Miller (Solanaceae)

Melia azedarach L. (Meliacaee)

Mirabilis jalapa L. (Nyctaginaceae)

Narcissus tazetta L. (Amaryllidaceae)

Nerium oleander L. (Apocynaceae)

Oxalis corniculata L. Oxalis pes-caprae L. (Oxalidaceae)

Papaver somniferum L. (Papaveraceae)

Parthenium hysterophorus L. (Asteraceae)

Table 1 Cont’d)

Erect perennial herb bearing small, whitish flower heads in groups

Erect annual herb bearing pretty red or white flowers

Small, delicate herbs bearing trifoliate leaves and yellow flowers.

Erect shrub bearing attractive pink or white flowers

Bulbiferous herb bearing white-yellow flowers

Perennial herb bearing tubular, pink flowers open at afternoon

Tree bearing hanging berrylike fruits.

Suberect herb bearing the tomatoes

Oxalic acid and oxalates

Alkaloids morphine, narcotine, codeine, papverine, thebaine

Entire plant

Latex from unripe fruits and other parts

Parthenin and other phenolic acids

Cardiac glycosides of the cardenolide type.

Entire plant

Entire plant

Salivation, abdominal pain, vomiting, diarrhoea, dizziness, central paralysis and collapse.

Alkaloid narcissine

Entire plant, particularly the bulbs

Common on roadsides and fallow land/

Cultivated as ornamental in most parts of the country.

Narcotic state, extremely slow respiration, pin-point pupils, death due to respiratory failure. Contact dermatitis, severe itching, eczema, photosensitivity.

First one is a vary common herb; the other is cultivated as ornamental.

Cultivated as ornamental, also found wild on stream sides.

Often cultivated for its flowers.

Commonly cultivated and found as an escape.

Cultivated in the plains and foothills.

Widely cultivated.

Hypocalcaemia, trembling, staggering. Prolonged use of small quantities causing deposition of calcium-oxalate in the kidney.

Severe gastroenteritis, with dizziness, high pulse rate, weak heart beat, unconsciousness, coma and death.

Consumption of the plant parts causes gastroenteritis

Alkaloid trigonelline

Entire plant

Vomiting, constipation or diarrhoea, breathing difficulty, cardiac weakening, CNS disturbance, paralysis.

Irritation in the throat, headache, vomiting, severe diarrhea, in some cases fever and circulatory collapse.

A resinous poison

Steroidal glycoalkaloids like solanine and others

Fruits

Green parts

Poisonous Plants of Islamabad

133

Safeda, Blue Gum

Chatri Dhodak Sun Spurge

Poinsettia

Morning Glory

Bhaiker, Basaka, Malabar Nut

Panch Phuli, Wild Sage

Chaora Sanatha, Privet

Mochni, Darnel

Eucalyptus globulus Labille (Myrtaceae)

Euphorbia helioscopia L. (Euphorbiaceae)

Euphorbia pulcherrima Willd. (Euphorbiaceae)

Ipomoea purpurea (L). Roth (Convolvulaceae))

Justicia adhatoda L. (Acanthaceae)

Lantana camara L. (Verbenacaee)

Ligustrum lucidum Ait. (Oleaceae)

Lolium temulentum L. (Poaceae)

Table 1 Cont’d)

Erect grass bearing green spikes on a straight axis

Shrub or small tree bearing small white flowers

Large bush bearing multicoloured, flowers and berry-like fruit

Undershrub bearing white, 2-lipped, flowers.

Climber with purple, bellshaped flowers

Shrub bearing red petal-like bracts

Erect herb with umbrella-like upper leaves

Tall with light grey trunk

Glycoside ligustrin

Alkaloids temuline, temulentine and lolin, and fungus infection may be involved

Seeds

Pentacyclic triterpenes, lantadene A and B

Alkaloid vasicine and essential oils

D-lysergic acid

Water-soluble caoutchouc.

Diterpene 12Deoxypharbol

Eucalyptol

All parts, especially the berries

fruits especially when unripe

Entire plant

Seeds

Leaves and latex

Latex

Oil of the plant

A wild grass

Contaminated flour cause trouble in GIS, CNS, cardiac weakness.

Saleem Ahmad Cont’d ........

Grown as a hedge plant.

Cultivated/ selfgrown in most parts of the country.

Common in subHimalayan tract, upto 4000'

Cultivated and found as an escape.

Cultivated as an ornamental.

Common on roadsides and fallow land.

Cultivated on roadsides and parks.

Gastric irritation, vomiting, purging, death in severe cases; leaves can cause severe dermatitis.

Gastroenteritis, diarrhoea and failure of blood circulation, photosensitivity may also occur.

Fall of blood pressure followed by a rise to normal, increase in heart beats amplitude and slow rhythm.

Disorientation, blurred vision, hallucinations, permanent psychological disturbances, death

Vomiting, purgation, delirium, externally blistering of the skin.

Burning of the mouth, oesophagus and stomach, vomiting, convulsions, oedema of lung, coma, death.

Epigastric burning, vomiting, dizziness, convulsions, can be fatal.

134

Kabikaj, Water Crowfoot

Arind, Castor Oil Plant

Pahari Shisham, Chinese Tallow Tree

Milk Thistle

Winter Cherry

Japanese Pagoda Tree

Peela Ganira, Yellow Oleander

Ban Okra, Chota Gokhru, Cocklebur

Ranunculcus sceleratus L. (Ranunculaceae)

Ricinus communis (Euphorbiaceae)

Sapium sebiferum Roxb. (Euphorbiaceae)

Silybum marianum (L.) Gaert. (Asteraceae)

Solanum pseudocapsicum L. (Solanaceae)

Sophora japonica L. (Papilionaceae)

Thevetia peruviana (Pers.) Schum. (Apocynaceae)

Xanthium strumarium L. (Asteraceae)

Table 1 Cont’d)

Branched herb bearing small bristle- covered fruits

Small tree bearing yellow flowers and peculiar fruit

Small tree with pinnate leaves and fruiting pods

Bush-like perennial herb bearing red berries.

Spiny herb with variegated leaves and spiny purple flowers

Tree with Shisham-like leaves and rounded fruits

Tall shrub bearing small flowers and dry bristly fruits

Annual herb bearing small yellow flowers

Carboxytractyloside

Entire plant when fresh and seeds

Alkaloid solanocapsine

Berries

Glycosides cerebrin, neriifolin, thevetin

Nitrates

Entire plant

All parts

Diterpene ester of phorbol.

Latex

Alkaloids cytisine, anagyrine

Toxalbumin ricin.

Seed coat.

All parts, especially seeds.

Glycoside ranunculin

Entire plant

Diuresis, sedation, pollen causes asthma, sinusitis and dermatitis in susceptible persons.

Burning mouth pain, numb tongue, vomiting, dilated pupils, heart block, collapse, death due to peripheral circulatory failure.

Stimulation of the respiratory centres, delirium, prolonged sleep; much dangerous to children

Retardation of cardiac impulse and sinus arrhythmias

Severe gastroenteritis, due to the reduction of nitrates to the more highly toxic nitrites

Skin contact of latex causes irruption and vesication; also said to be carcinogenic.

Severe GIT irritation, vomiting, bloody diarrhoea, cyanosis, convulsion, circulatory collapse and death.

Dermatitis, severe gastroenteritis, mouth and lips inflamed.

Common in disturbed and unused places.

Cultivated on roadsides and parks.

Sometimes cultivated in parks.

Cultivated as an ornamental.

Found in fallow land.

Cultivated in parks and roadsides as an ornamental tree.

Cultivated and escaped.

Common in wet places.

Poisonous Plants of Islamabad

135

136 impenetrable thickets in waste places. Its ripe fruits are harmless and are eaten by people and also used in herbal medicine, but when unripe they can cause poisoning. Oleander (Nerium oleander) is quite common in parks and roadsides as it has quite beautiful floral display, but contains deadly toxins. Blood weed (Asclepias curassavica), Madagasar periwinkle (Catharanthus roseus), night jessamine (Cestrum nocturnum), Lakspur (Consolida ambigua), golden dewdrop (Duranta repens), Poinsettia (Euphorbia pulcherrima), morning glory (Ipomoea purpurea), Privet (Ligustrum lucidum), four O’clock (Mirabilis jalapa), opium poppy (Papaver somniferum) are commonly cultivated in parks and gardens. The harm caused by these poisonous plants is often not serious, and is primarily restricted to gastrointestinal irritation or mild nervous system effects, which are usually cured by a physician. However, there have been cases of death resulting from consumption of parts of highly poisonous plants. For example, several young people have died in Islamabad after eating the odd looking fruit of the yellow oleander (Thevetia peruviana), leaves of night jessamine (Cestrum nocturnum) and oleander (Nerium oleander). Use of the narcotic Hemp (Cannabis sativa), which is common in this country, has led to death of a few addicts due to overdose (Chopra et al., 1984). Some of the poisonous plants like Butea, Calotropis, Centella, Datura, Solanum and Withania are used in herbal medicines in Pakistan (Nasir and Ali, 1970). The use of incorrectly prepared herbal medicines also cause poisoning, especially among rural people. Preparations for enhancing sexual performance are also commonly sold, which often contain extracts from toxic plants, animals constituents, and various minerals that cause harm to the uniformed users. Green vegetative parts of commonly used “vegetables”, like potatoes (tubers) and tomatoes (berries) can lead to poisoning since few people are aware of their toxicity. Of the various plant parts, the attractive-looking fruits of cultivated or wild plants cause poisoning more frequently and victims are often children. Fruits of black nightshade (Solanum nigrum) locally called ‘Makoh’ is a common roadside plant of Islamabad. Its ripe fruits are harmless and are often eaten by children. But its unripe fruits are toxic and if consumed in large numbers can be very harmful. Datura species are often seen in groups and are attractive because of their large, funnel-shaped white flowers. Its prickly fruits contain a large number of seeds that are used by

Saleem Ahmad

Hakims in anti-asthma preparations. But used alone, the seeds can cause poisoning with hallucinogenic effects or death. Leaves and/or stems of some plants are also consumed out of curiosity, which may cause poisoning (e.g. the stem of the commonly cultivated Diffenbachia causes severe inflammation if it comes in contact with the mouth). It is important that people abstain from consuming any unconventional plant parts, even if they may not appear harmful. Children at home and in parks should be kept under watch and trained not to taste plant parts. In the event of ingestion and appearance of any undesirable symptom, the child should be taken to a hospital. As a preveitive measure, it is imperative that public awareness about the poisonous plants in ones vicinity be emphasized to prevent any unnecessary or even tragic events associated with local poisonous plants.

References Ahmad, S., Khattak, Z. 2001. Quantitative studies on the vegetation of Islamabad. Pakistan Journal of Scientific and Industrial Research, 44: 279-285. Anderson, E. 1967. Plants, Man and Life, pp. 124-135, University of California Press, Berkley, USA. Baquar, S.R.H. 1989. Medicinal and Poisonous Plants of Pakistan, 506 pp., Printas Karachi, Pakistan. Baquar, S.R., Tasnif, M. 1967. Medicinal Plants of Southern West Pakistan, pp. 7-119, Pakistan Council of Scientific and Industrial Research, Karachi, Pakistan. Champion, H.G., Seth, S.K., Khattak, G.M. 1965. Forest Types of Pakistan, pp.130-140, Pakistan Forest Institute, Peshawar, Pakistan. Chopra, R.N., Badhwar, R.L., Ghosh, S. 1984. Poisonous Plants of India, vol.1, pp. 13-23, Academic Publishers, Jaipur, India. Chopra, R.N., Nayar, S.L. Chopra, I.C. 1956. Glossary of Indian Medicinal Plants, pp. 13-63, Council of Scientific and Industrial Research, New Delhi, India. Connor, H.E. 1997. The Poisonous Plants in New Zealand. E.C. Keating (ed), pp. 14-27, 2nd edition. Government Printer, Wellignton, New Zealand. Cooper, M.R., Johnson, A.W. 1984. Poisonous Plants in Britain and their Effects on Animals and Man. pp. 130-305. Her Majesty’s Stationery Office, London, UK. Dimitrios, B. 2006. Sources of natural phenolic antioxidants. Trends in Food Science & Technology. 17: 505-512.

Poisonous Plants of Islamabad

Dymock, W., Warden, C.J.H., Hooper, D. 1890-1893. Pharmacographia Indica. vol. 1, pp. 59-81, Education Society’s Press, Bombay, India. Frohne, D. Pfander, H.J. 1984. A Colour Atlas of Poisonous Plants, pp. 113-291, Wolfe Publishing Ltd, London, UK. Grieve, M.1979. A Modern Herbal. C.F. Level (ed.), pp. 170-171, Jonathan Cape, London, UK. Heywood, V.H. Chant, S.R. 1982. Popular Encyclopedia of Plants, pp. 116-117, Cambridge University Press, UK. Hooker, J.D. (ed.) 1875-1888. The Flora of British India. vols. 1, pp. 20-21, L. Reeve & Co. Ltd. Ashford, Kent, UK. Ikram, M., Hussain, S.F. 1978. Compendium of Medicinal Plants. 167 pp., Pakistan Council for Scientific and Industrial Research, Peshawar, Pakistan. Kingsbury, J.M. 1964. Poisonous Plants of the United States and Canada. pp. 9-28, Prentice Hall, Englewood Cliffs, New Jersey, USA. Lewis, W.H., Elvin-Lewis, M.P.F. 1977. Medical Botany: Plants Affecting Man’s Health. pp. 11-62, John Wiley & Sons, New York, USA. Modi, N.J. 1945. Textbook of Medical Jurisprudence

137 and Toxicology (First edition, 1920). Tripathy Ltd., Bombay, India. Nasir, E., Ali, S.I., (eds.). 1970. Flora of West Pakistan/ Nos. 1-190, PARC, National Herbarium, Islamabad and Department of Botany, University of Karachi, Pakistan. Pandey, B.P. 1984. Economic Botany. 476 pp., S. Chand & Company Ltd., New Delhi, India. Polunin, O., Stainton, A. 1985. Flowers of the Himalaya. vol. 1, pp. 11-391, Oxford University Press. Delhi, India. Stewart, R.R. 1972. An Annotated Catalogue of the Vascular Plants of Pakistan and Kashmir. 1028 pp., Fakhri Printing Press, Karachi, Pakistan. Stewart, R.R. 1957. The Flora of Rawalpindi District. Pakistan Journal of Forestry, 7: 240-300. Stuart, M. 1979. The Encyclopedia of Herbs and Herbalism. 283 pp., Orbis Publishing Ltd., London, UK. Tampion, J. 1977. Dangerous Plants, pp. 73-81. David and Charles Publication, Newton Abbot, London, UK. Willis, J.C., Airy Shaw, H.K. 1985. A Dictionary of the Flowering Plants and Ferns. 1245 pp., 8th edition, Cambridge University Press, UK.

Pak. j. sci. ind. res. Ser. B: biol. sci. 2012 55 (3) 138-144

Effects of Passiflora foetida Linn. (Passifloraceae) on Genital Tract, Serum Estradiol, Pituitary Gonadotropin and Prolactin Level in Female Adult and Immature Ovariectomized Rats Bleu Gome Michel*a, Kouakou Koffi b, Toure Alassane c and Traore Flavien a a

Laboratory of Animal Physiology, UFR Biosciences, University of Cocody, Abidjan, 22 BP 582 Abidjan 22, Côte d’Ivoire b Laboratory of Biology of Reproduction and Endocrinology, UFR Biosciences, University of Cocody, Abidjan, 22 BP 582 Abidjan 22, Côte d’Ivoire c Division of Anatomy-Pathology, Central Laboratory of Veterinary Medicine (LANADA), Bingerville, BP 206 Bingerville, Côte d’Ivoire (received August 26, 2011; revised August 6, 2012; accepted September 4, 2012) Abstract. The effects of the extracts of Passiflora foetida on the genital tract, serum estradiol, pituitary gonadotropin (LH and FSH) and prolactin were studied in female adult (120-140 g) and immature ovariectomized (30-40 g) rats. Results showed that the aqueous extract increased significantly both the ovary and uterus weight whereas the hexane extract increased the weight of the ovary only and the methanol extract increased the weight of uterus only. Histological examination of these organs indicated that P. foetida treated rats were in estrous or proestrous phases of the estrous cycle. The hormone analysis showed that the serum LH was significantly increased by 17b estradiol and by the three extracts dose dependently in immature ovariectomized rats. Moreover, the aqueous extract increased significantly serum estradiol and pituitary gonadotropins and prolactin in adult non ovariectomized rats. Keywords: Passiflora foetida, ovary, uterus, estradiol, gonadotropins, prolactin

Introduction

tuents of this plant are flavonoids, glycosides, phenolic compounds, fats, steroids and alkaloids (Patel et al., 2011; Ingale and Hivrale, 2010; Echeverri et al., 2001; Cambie and Ash, 1994). This study was aimed to evaluate the effects of Passiflora foetida leaves on the genital tract, estrogen and pituitary hormone levels in female wistar rats.

According to the World Health Organization, 80% of the population in Africa depends on traditional medicine for their health-care needs (WHO, 2002). This continent is endowed with a rich biodiversity estimated about 40,000 plant species (Mahunnah, 2002) and more than 4000 species are used medicinally (Bosh et al., 2002). Infertility is one of the most crucial health problems which are treated in the African traditional medicine. This reproductive problem causes divorces, instability and polygamy in many families where some women considered as sterile are victims of violence (Papreen et al., 2000; Unissa, 1999). Hence, most of them use medicinal plants to recover their fecundity (KambuKabangu, 1990) due to poverty. Passiflora foetida is a medicinal plant used by population in the south of Côte d’Ivoire to cure infertility in women (N’Guessan, 2008; 1995). Furthermore, it serves to treat other diseases such as snakebites, epilepsy, and abscess (N’Guessan, 2008; 1995; Adjanohoun and Aké-Assi, 1979). It is used in Asia continent as emmenagogue and as remedy for asthma, biliousness, hysteria, giddiness and headache (Dhawan et al., 2004). The major phytochemical consti-

Materials and Methods Plant material. The leaves of P. foetida were collected from the north and the south of Abidjan, the economic capital city of Côte d’Ivoire and authenticated in the Laboratory of Botany (UFR Biosciences) of the University of Cocody, Abidjan. A voucher specimen was deposited in the botanical garden of this University under the number 746B. Preparation of the extracts. The collected plant material was dried at an ambient temperature (30±2 °C) without exposure to sun light and crushed to obtain a powder which was divided into three parts, 50 g of each were macerated separately for 24 h in water (1500 mL), hexane (750 mL) and methanol 95° (750 mL), filtered using Whatman filter paper number 1 and concentrated in an air circulating oven at 50 °C until total dryness.

*Author for correspondence; E-mail: [email protected]

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Phytochemical Effects of Passiflora foetida on Rats

The aqueous, hexane and methanol extracts obtained (yield 20.23%, 6.83% and 28.03% respectively) were then stored at 4 °C in a refrigerator for the experimental studies. Effects on the genital tract. Adult female wistar albino rats (120-140 g) were divided into groups of 5 rats each and administered separately the dose of 500 mg/kg body weight of each extract for 14 and 28 days. The control group was treated with olive oil. All the treatments were given orally, using an intragastric cannula. At the day after the last treatment, the rats were weighed and sacrificed by decapitation under light ether anesthesia. The ovary and uterus of each rat were dissected out and weighed rapidly using a sensitive balance. The adrenal and kidney were also dissected and weighed. Additionally, the dissected organs were fixed in bouin’s fluid for 24 h, dehydrated and embedded in paraffin. The paraffin sections were cut at 4 µm and stained with haematoxylin-eosin for histological examinations. Effects on LH level of immature ovariectomized rats. The immature female albino rats (30-40 g) were bilaterally ovariectomized (ovx) under light ether anesthesia through lateral incision in the skin just below the last rib (Keshri et al., 1995; Zarrow et al., 1964). After a post operative rest period of 7 days, the rats were divided into 8 groups of 6 animals each and treated separately with olive oil (control group), 0.02 mg/kg body weight of 17b estradiol, the high dose (500 mg/kg body weight) and low dose (250 mg/kg body weight) of each extract. All the treatments were given orally for 7 consecutive days and 24 h after the last treatment, the rats were anesthetized with ether and sacrificed by

decapitation. The blood samples were collected into non heparinized tubes and centrifuged at 2580 rpm for 10 min. The serum was then separated and stored at -20 °C until LH analysis by the ELFA technique (Enzyme Linked Fluorescent Assay) using specific kit (BioMerieux, Lyon, France). Effect on serum estrogen, gonadotropins and prolactin of adult rat. The aqueous extract was found to be the most active extract on the genital tract and on LH level of ovx rat. Hence, it was used to determine the effect of P. foetida on estrogen and pituitary hormones. For this experiment, female adult rats were divided into two groups of 5 rats each. The first group served as control and received olive oil only and the second group was treated with a dose of 500 mg/kg body weight of the aqueous extract by gavage for 28 consecutive days. At the end of the treatment, the rats were sacrificed under ether anesthesia and the collected blood samples were analyzed for the level of estrogen (E2), gonadotropin (LH and FSH) and prolactin (PRL) in the serum by the same methods used in the previous experiment. Statistical analysis. The data were expressed as mean±SEM. Statistical analysis of the variance between control and experimental values were done using student’s t-test. A probability level of less than 5% (p