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Open Access
Molecular characterization of piper species usiing RAPD technique
(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 2002) Murugan, C; KAU; Sujatha, V S
The study on "Molecular characterization of Piper species using RAPD techniques" was conducted in the Department of Plantation Crops and Spices and CPBMB, College of Horticulture, Vellanikkara, Thrissur from December, 1999 to August, 2001. Material included 13 species/accessions of the genus Piper including Piper nigrutn Linn. These were analysed with 20 selected oligomer decamer primers using PCR technique and agarose gel electrophoresis. The selection of ideal DNA isolation technique was done based on observations of purity of bands, quality of the DNA bands and quantity of DNA recovered. The protocol of Edwards et al. (1991) was selected for DNA isolation with additional step of chloroform-isoamyl alcohol treatment. Fifty decamer primers were screened using DNA. of P. nigrum and 20 were selected based on the number of bands produced, reproducibility of the bands and distinctness of the RAPD profile. RAPD profile of the 13 species of Piper were compared for genetic similarity index (using Jaccard's coefficient), with all 20 selected primers separately. Pooled similarity of the twenty primers put together was found out for 9 species which gave amplification for all the 20 selected primers. The results of pooled analysis was subjected to cluster analysis (SAl-IN) and phenetic dcndrogram was constructed employing UPGMA. Four clusters were obtained, comprising of two accessions of P. nigrum In the first, two accessions of P. longum in the second, P. colubrinutn and P. attenuatum forming the third cluster and P. chaba, P. belle and P. arboreum forming the fourth.
Open Access
Character association of seeds on plant morphology in snakegourd (trichosanthes anguina L.)
(Department of Olericulture, College of Horticulture, Vellanikkara, 2000) Ashok, P; KAU; Rajan, S
The research project 'Character association of seeds on plant morphology in snakegourd (Trichosanthes anguina L.)' was carried out in College of Horticulture, Kerala Agricultural University, Vellanikkara, Thrissur during the period 1999-2000. The study envisaged to find out the association of various seed characters on the vegetative characters of snakegourd, classification of accessions based on the seed index, growth index and yield index as obtained from principal component analysis and to formulate keys for varietal identification. The 25 accessions were catalogued based on the IBPGR descriptor list. Significant differences for various seed characters, growth characters and yield characters were noticed among the accessions. Correlation studies have shown that the number of seeds per fruit strongly associated with days to male flower opening and fruit maturity. Seed size index was strongly associated with leaf size. The highest seed index value for TAl 08, the highest growth index value for TA 118 and the highest yield index value for TA 114 were obtained as revealed from principal component analysis . . Twenty five accessions were grouped into different clusters based on the seedindex, growth index and yield index. Based upon the seed characters (seed size, seed coat colour and 100 seed weight), growth characters (leaf lobes, leaf pubescence and tendril length) and fruit characters (fruit skin colour, fruit girth, fruit length and fruit weight) keys were developed for varietal identification.
Open Access
Parasexual hybridization of piper nigrum and piper colubrinum through protoplast fusion
(Department of Plantation Crops And Spices,College of Horticulture, Vellanikkara, 2000) Resmi, Paul; KAU; Rajendran, P C
Black pepper is the most important export oriented commodity and foreign exchange earner among the Indian spices. Ravages due to diseases, particularly the most devastating Phytophthora foot rot caused by Phytophthora capsici is one of the major constraints in the production of black pepper all over the world. Piper colubrinum, a wild relative of black pepper is found to be immune to foot rot disease. Non – existence of cultivar level tolerance or resistance against foot rot disease in black pepper necessitated the incorporation of incompatible wild relatives through parasexual hybridization. This study was undertaken in the Department of Plantation Crops and Spices and the Plant Tissue Culture laboratory of the Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara between September 1998 to April 2000. The cultures of P. nigrum and P. colubrinum were initiated in test tubes and were maintained at 26 + 20C temperature and 60 to 80 per cent humidity. The green leaves excised from axenic cultures of both the species were treated with cell wall degrading enzymes, cellulose and pectinase maintained at proper osmotic concentration. In P. nigrum maximum yield was observed at 1.4 per cent cellulose and 0.34 per cent pectinase. Cellulase and pexctinase at a concentration of 1.0 per cent and 0.217 per cent respectively recorded highest yield in P. colubrinum. In both the species 0.6 M osmoticum was found to be optimum to maintain the osmotic potential of the isolation solution. Highest yield of protoplasts was recorded in both the species during 21 h of digestion. Filtration-centrifugation technique was found to be superior in purifying the Piper protoplasts compared to the sucrose floatation method. Centrifugation at 1000 rpm for three minutes was found to be best for purifying P. nigrum protoplasts. For purifying P. colubrimum protoplasts, 600 rpm for three minutes was found to be optimum. Highest viability was noticed at 0.55 M and 0.65 M osmoticum in 1.0 per cent cellulose and 0.28 per cent pectinase during 21 h of digestion in P. nigrum. In P. colubrinum, maximum viability was observed at 0.4 M osmoticum in the enzyme mixture 1.0 per cent Cellulase and 0.186 per cent Pectinase during 18 h of incubation. Protoplasts of both the species when cultured on modified MS medium formed no cell wall and have not undergone any division. A age of the cultures advanced, proptoplast viability decreased in P. nigrum and P. colubrinum. All the protoplasts died by the second week in both the species. The protoplasts of both the species were heterogenous in terms of size. Fusion of the protoplasts was not observed after PEG treatment in the present study.
Open Access
Standrdisation of population density in watermelon (Citrullus lanatus (Thunb.) Mansf.)
(Department of Horticulture, College of Agriculture, Vellayani, 2000) Bindukala, A R; KAU; Abdul Vahab, M
The present investigation on "Standardisation of population density in watermelon [Citrullus lanatus (Thimb.) mansf.]" was conducted at the Instructional Farm, College of Agriculture, Vellayani, Thiruvananthapuram during February - May, 1999 .. Three watermelon varieties viz., Sugar Baby, Arka Manik and MHW -6 were tried under six population densities to find out their direct and indirect effects on vegetative yield and quality attributes in watermelon. Majority of the characters were significantly influenced by the direct as well as interaction effects of the population density and varieties. Among the varieties, Arka Manik was earlier for male and female flower opening and for days' to first harvest. However, the node to first female flower was in Sugar Baby which also had more female flowers planr". The population density significantly influenced majority of the vegetative characters. Lower to moderate population density had shorter crop duration, branches planr ' and vine length. Different density levels significantly influenced the yield characters. Total fruits planr ', Total fruits plor ', Marketable fruits planr", 2 Marketable fruits plor ' ,Marketable yield plant"! were maximum in plants grown at highest density of 10000 plants ha". The variety Sugar Baby was first in all these yield characters. Sugar Baby also had the highest values for diameter, flesh thickness, rind thickness, fruit girth and seeds fruir '. The hybrid MHW-6 had maximum average fruit weight, fruit length and 100 seed weight. Among the quality parameters, Sugar Baby had maximum values for total sugars and reducing sugars whereas, Arka Manik recorded the highest values for total soluble solids, non-reducing sugars and iron content. The plants at moderate spacings recorded highest values for the quality characters. Majority of the characters were influenced by the interaction of population densities and varieties also. Sugar Baby at a moderate population density of 3333 plants ha! had maximum branches plant"! .. The number of fruits plor ' was highest in Sugar Baby at the highest population of 10000 plants ha:". This treatment combination also had maximum marketable yield plor ' registering maximum yield of 55 t ha! and maximum income of Rs. 165000 with the highest bene:{it cost ratio of 3.00 : 1.00.
Open Access
Biodiversity of medicinal plants in oil palm plantations
(Department of Horticulture, College of Agriculture,vellayani, 2000) Sarada, S; KAU; Sreekandan Nair, G
A study on 'Biodiversity of medicinal plants In oil palm plantations' was carried out at the oil palm plantations, Kulathupuzha of the Oil palm India Ltd., Kollam district from Januarv 1998 to ., January 1999. The objectives of the study were to identify the medicinal plants in the oil palm plantations, to study the growth behaviour of selected medicinal plants and to assess the inf1uence of shade on pharmacologi.cally active constituents. A total of 80 sampling units were taken us rug stratified random sampling technique, the strata being young, medium and mature oil palm plantations and open conditions. The medicinal plants In the inter spaces of young, medium and mature oil palm plantations and in the open were identified and quantified by random sampling technique using 1.0m2 frame. A total of 85 plants species were identified in the four different strata belonging to 79 genera and 36 families. No ne of the plants were endemic. There were 74 indigenous and 10 exotic/naturalized plants Ten important medicinal plant species were selected for detailed study and there growth behaviour was monitored for one year. They were Chro mol aena odorata, Cyclea p elt at a, E l e p h a nt o p u s scaber, Em if i a so nchifol ia, H emidesmus in dicu s, Hypt is s u av eol ens, Phyll antlius am arus, Si d a rh omb ifol i a, So/anum m el o ngena var. insariurn and Vernonia cinerea. Ch rys op o gon acicul at us dominates all the four strata \\.j b high relative density and frequency. Cyrt o cocc um trigonum is another dominating species with high relative density in young and 0):11 re oil palm plantations. Nar egami a a/ala is a dominant and abundant species occurnng in mature plantation. He mi d e s nius in die us is o cc urr i ng f re que n t I y in all the f 0 u r s t r a t a. E! e ph a nt 0 p 11 S sea b e roe cur s fr e ,- u e nt 1 v m young plantation and Chromolaena odorata in medium plantation. Phyll anth us amarus is more frequent in mature plantation and open conditions. H olo st emma adakodien also occur frequently in mature plantation. Young oil palm plantation and open conditions were found to be the most similar strata with more number of species m common. Mature oil palm plantations and open conditions were found to be the most dissimilar strata in vegetation pair wise analysis. Medi urn oil palm plantation was found to have higher concentration of dominance as expressed by Simpso n ' s Index. Abundant species occurs more in mature oil palm plantation and very abundant species occurs more in medium oil palm plantation. Evenness index was maximum in open conditions. Growth characters like plant height, number of branches, plant spread, height of first branch, number of leaves, inter nodal length and stem girth showed the lanky growth of the selected ten medical plants in medium and mature oil palm plantations which were more shaded compared to young plantation and open conditions. Fresh and dry weight of officinal part was more In young plantation and open conditions compared to medium and mature oil palm plantations. Higher biomass production was also obtained in young oil palm plantation compared to medium and mature plantations. The amount of essential oil in the roots of He m id esm us ind i c us was highest under open conditions and lowest in mature oil palm plantation. There was no significant difference between the amount of essential oil in the roots under young and medium oil palm plantations. • . I, The solasodine content in the fruits of Solanum mel ongena var. i ns anum was highest under open conditions and lowest in medium oil palm plantation. There was no significant difference between the solasodine content in fruits in young plantation and open conditions; medi urn plantation and mature plantation. The solasodine content in roots was also highest under open conditions and lowest is mature plantation. There was no significant difference between the so lasodine content in roots in young plantation and open conditions; medium plantation and mature plantation.
Open Access
Comparative biology and growth behaviour of Pennisetum polystachyon (L.) Shult and P. pedicellatum Trin
(Department of Agronomy, College of Horticulture, Vellanikkara, 2000) Karthik, A; KAU; George Thomas, C
Pennisetum polystachyon (L.) Shult. and P. pedicellatum Trin. are two major grass weeds that are widespread in young plantation crops and non-arable areas of the tropics. The present investigation was undertaken to have an understanding on the biology, growth behaviour and fodder production potential of these two Pennisetum spp. at the Research Farm, College of Horticulture, Vellanikkara during 1998-2000. Germination tests were conducted at monthly intervals. P. polystachyon did not show any dormancy and started germination as soon as the seeds are collected (December). P. pedicellatum seeds remained dormant during December and January and it started germination by February only. Both species showed above 80 per cent germination by late March. Germination percentage started to decline in P. polystachyon after April. Phenological characters of both species were noted from a natural population during the year 1999. Seeds of Pennisetum spp. started germination soon after the receipt of monsoon showers by the end of April. Flowering started after the completion of the rainy period in September. Plants started senescence by mid December in P. polystachyon and mid November in P. pedicellatum. Both Pennisetum species studied showed remarkable regeneration ability from different cutting heights. The percentage of plants regenerated was similar in both species, Allelopathic reactions of roots, leaves and seeds were also studied. Boiled and unboiled extracts were prepared and germination test were conducted using cowpea, bhindi and cucumber as test crops. The cold and boiled leaf extracts of both species did not show any effect on germination of test crops. However, cold root extract of both Pennisetum spp. reduced the germination percentge of bhindi. Cold root extract of both species also showed some promotery effect on shoot length in cowpea. Seed extracts showed inhibitory effects only on cowpea germination. Growth was comparatively fast in P: pedicellatum than P. polystachyon during its early stages. P. polystachyon showed higher leaf : stem ratio than P. pedicellatum. Growth indices like LAR, LWR, RGR, AGR and NAR were similar in both Pennisetum spp. In terms of total green fodder production, P. pedicellatum was superior to P. polystachyon. Two cuttings, one at 70 days after sowing and the other just before flowering recorded maximum fresh weight of fodder. Though crude protein content was similar in both the species, P. pedicellatum showed a higher crude fibre content than P. polystachyon. The results indicate that as the fodder quality is good, utilizing these grasses as cattle feed can be one of the strategies for managing them. However, cultivation of these grasses for fodder purposes involves some risks because of their high reproductive capacity and persistence.
Open Access
Genetic transformation and hairy root culture in ada-kodien
(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 2001) Karmarkar, Shirish Hari; KAU; Keshavachandran, R
The present study was undertaken at the Department of Plantation Crops and Spices, College of Horticulture, Kerala Agriculture University, Vellanikkara during May 1999 to October 2000 The study was undertaken to standardize the procedure to genetically transform Holostemma ada-kodien and induce hairy roots it was also envisaged to standardise the biochemical techniques for the estimation of secondary metabolites in the roots of Holostemma. Hairy roots were induced by infection of Holostemma explants with a gram negative soil bacterium, Agrobacterium rhizogenes. Leaf segments, shoot buds, intermodal segments, seedling hypocotyls and callus were used as explants for hairy root induction. Among them, the seedling hypocotyls showed highest potential for hairy root induction followed by shoot buds. Leaf segments, intermodal segments and callus did not induce hairy roots. Different strains of Agrobacterium rhizogenes viz PcA4, 15834, A4, 8196 and 2659 were evaluated for their ability to induce hairy roots in Holostemma explants. The strain PcA4 showed the highest potential for hairy root induction, followed by strains 15834 and A4. The strains 8196 and 2659 did not induce hairy roots. Direct inoculation of bacteria on wounds induced hairy roots on seedling hypocotyls only. Co-culture of wounded explants with bacteria induced hairy roots on seedling hypocotyls and shoot buds. In the direct inoculation method, the nature of bacterial inoculum and the intensity of bacterial inoculum applied on wounds influenced the transformation. Bacterial cell suspensions when applied on wounds induced transformation. Less intensity of bacterial inoculum when applied on wounds gave greater transformation frequencies and vice versa. In the co-culture method, the intensity of bacteria present during co-culture, the co-culture time and the shaker speed influenced the transformation. Comparative evaluation of varying intensity of bacterial population during co-culture and the co-culture time showed that, both less intensity of bacterial inoculum with more co-culture time and vice versa showed almost similar transformation frequencies (14.40 % and 14.51 % respectively). The shaker speed of 100 rpm gave the highest transformation percentage than 50 rpm speed. Shaker speed of 150 rpm did not induce any transformation. Application of different concentration of NAA prior to direct inoculation of bacteria on wounded explants did not aid in transformation. Addition of 2 mg 1-1 NAA in the co-culture medium, however, increased the transformation frequency from 25 per cent to 75 per cent. Photoperiod influenced the transformation frequencies. A photoperiod of 16 h light was found to be the best for hairy root induction in Holostemma. Media influenced the hairy root induction. Full strength MS medium favoured hairy root induction while ½ strength MS medium did not favour hairy root induction. Hairy roots were induced in a period of one to four weeks in all the treatments. The induced hairy roots showed altered phenotypes. The hairy root obtained directly from explants without NAA treatment were whitish, hairy and showed negative geotropism. The hairy roots obtained after NAA treatment were brownish yellow and were induced from calli formed on wounds after infection. Hairy roots obtained on infection with strains PcA4, 15834 and A4 showed the presence of agropine confirming their transformed nature. Normal roots did not show presence of opine (s). Hairy roots showed greater sensitivity to lower concentrations of NAA than the normal roots. At 10-9 M concentration of NAA the hairy roots showed lateral branch formation. The tubers and in vitro induced callus of Holostemma were tested for the presence of amino acids, essential oils, triterpenoids and sterols. Six amino acids were found in the root tubers while an additional amino acid (Rf 0.48) that may be L Proline or Cysteine MHC was found in the callus. Root tubers and callus showed the presence of essential oils, triterpenoids and sterols. Root tubers showed the presence of more number of essential oils and triterpenoids than the callus. The callus showed the presence of new terpene compounds. The two sterols present in root tubers and calli were identified to be α –amyrin and β –amyrin
Open Access
Evaluation of genetic stock of sanghupushpam (Clitoria ternatea L.) for yield, Alkaloid content and nitrogen fixing potential
(Department of Horticulture, College of Agriculture, Vellayani, 2000) Deepa, S Nair; KAU; Reghunath, B R
The present study entitled “Evaluation of genetic stock of ‘Sanghupushpam’ (Clitoria ternatea L.) for yield, alkaloid content and nitrogen fixing potential” was carried out at the Department of Horticulture, College of Agriculture, Vellayani from June 1999 to January 2000. Seeds of twenty different accessions were collected from various locations from inside and outside the state. Thirteen accessions having high rate of seed germination were raised as intercrop in young coconut garden and maintained till seed maturation stage. The performance of the accessions in terms of growth, yield and physiological parameters were evaluated. Growth and yield parameters with respect to shoot, pod, root and root nodule characters were evaluated. Physiological parameters evaluated included leaf area index, leaf area duration, net assimilation rate, crop growth rate, relative growth rate, absolute growth rate and harvest index. Number of effective nodules was taken as an index for assessing nitrogen fixing potential of the plant. Leaf yield, shoot yield and root yield were significantly superior in the accession MP-90. MP-83 recorded significantly superior pod yield. The number of nodules was the highest in accessions MP-76 and MP-82. Crude alkaloid content was significantly superior in seeds of MP-74 and MP-76. Six accessions were selected based on yield , nodule characters and crude alkaloid content viz ., MP-90, MP-74, MP-83, MP-78, MP-76 and MP-82. Results of the present study indicated that Clitoria ternatea, is not a good proposition as an intercrop in young coconut garden. However, it may be worth studying the performance of the crop as a pure crop under open condition or as an intercrop in coconut gcirden with comparatively lesser shade (less than 50 per cent) than the situation of the present study.
Open Access
Selection index and activation of seedling growth in mangosteen
(Department of Pomology and Floriculture, College of Horticulture, Vellanikkara, 2002) Mohamed, Yosuf L; KAU; Sajan, Kurian
The experiments on "Selection index and activation of seedling growth in Mangosteen (Garcinia mangostana L.) were conducted in the central orchard attached to the Department of Pomology and Floriculture of the College of Horticulture from March 2000 to February 2002 with the broad aims of standardizing selection indices as well as activation of the seedling growth in mangosteen. Two separate experiments, one approach envisaging growth regulators and the .other involving arbuscular mycorrhizal fungi (Glomus mosseae and Glomus fasciculatum and Azospirillum with single super phosphate were taken up. The important results are as follows. Analysis of leaf biochemical characters of the mother plants belonging to different age groups «25, 25-50, 51-75 and >75 years) revealed that nitrogen, phosphorus, potassium and crude protein, were highest in the lowest «25 years) age group and least values were in the highest (>75 years) age group, except sodium, total phenols and total carbohydrates, which were maximum in the age group of 51-75 years. The number of seedless fruits and almost all the morphological characters of the fruit were highest in the age group of 51-75 years. Number of one seeded, two seeded and three and more than three seeded fruits were also varying among the four age groups. All seed morphological characters and number of ungerrninated seeds showed maximum values in the age group of 51-75 years and >75 years. Seeds obtained from the age group of 51-75 years showed earliness in germination but the percentage of germination was highest in the age group of 25-50 years. The biochemical characters of seed revealed that nitrogen, phosphorus, potassium, crude protein, total carbohydrates, total sugar and total phenols were maximum in the age group of25-50 years, whereas sodium and abscisic acid were highest in the age groups of less than 25 and more than 75 years, respectively. The morphological characters of seedling such as, height, girth, total number of leaves per seedling, total leaf area, survival rate at twelve months stage and root characters were highest in 25-50 years age group whereas the root and shoot dry weight were maximum in the less than 25 years group. Nitrogen, phosphorus, potassium, crude protein and sodium levels in the leaves of the seedling were highest in the age group of less than 25 years. Chlorophyll a, b, total phenols, total carbohydrates and abscisic acid contents were higher in the age group of 25-50 years. Maximum fruit index was observed in the age group of 51-75 years but seed and seedling index were highest in 25-50 years age group. Most of the seed characters were positively correlated with seedling characters. Though all the fruit characters were positively correlated with seed characters they were negatively correlated with seedling characters. Fruit index, seed index and seedling index were separately predicted using the prediction models fitted by multiple regression equations of principle components one and two with corresponding adjusted R2 values. Dominant characters among the age of mother plant, fruit, seed and seedling characters separately were found out by figurative plot principal component analysis based on morphological data of the above characters individually. Using step wise regression major characters contributing to seedling index were identified as age of the mother plant, seed thickness at centre, total leaf area, survival rate, fresh weight of root and plant, root length and total number of roots, which contributed maximum variation to seedling morphological characters and prediction model for fruit index, seed index and seedling index were fitted. To establish the similarities (or dissimilarities) between the four age groups, hierarchical cluster analysis using euclidean distance was performed which revealed that the younger two age groups (less than 25 years and 25-50 years) of the mother plants were the closest groups with respect to all characters. The four using discriminant functions were fitted to discriminate the four age groups with corresponding D2 values. The criteria and criterion were determined to describe the age groups of selected fruit, seed and seedlings collectively. By giving equal importance to age of the mother plant, fruit index, seed index and seedling index the selection index was worked out and the best age group of the mother plant for obtaining maximum growth of seedlings was identified as 25-50 years. Based on the correlations between various characters and growth index, the above or below average values with respect to each of the attribute (important and visual) belonging to the fruit, seed and seedling characters were identified. The mid values of the each attribute in the best age group were fixed as the index for selection. Among the growth regulator sprays for activating the slow growth in mangosteen, the best treatments were GA 150 mg r' followed by IAA 250 mg r'. Biochemical studies on the growth regulators treated plants revealed that the treatment that showed maximum growth also recorded the highest values of nitrogen, phosphorus, potassium, crude protein, chlorophyll a, b, total chlorophyll, total phenols, total carbohydrates and abscisic acid content. The treatment, which showed intermediate growth recorded intermediate values except for sodium, where it was maximum. The least content was observed in the treatments with least growth. The control plant showed intermediate values between the. values of plant with intermediate and least growth. Best treatment for activating the seedling growth among the treatments with arbuscular mycorrhizal fungi and Azospirillum inoculations were identified as, the combinations of Glomus Jasciculatum 5 g + Azospirillum 109 + single super phosphate 109 followed by GlomusJasciculatum 5 g + Azospirillum 20 g + single super phosphate 109. A rhythmic pattern was observed with the treatments showing best seedling growth also recording higher values of nitrogen, phosphorus, potassium, crude protein, chlorophyll a, b, total chlorophyll, total phenol total carbohydrates and the abscisic acid content, and the treatments showing in intermediate growth recording intermediate values except sodium. The least values were in treatments that showed least growth. The control plant showed intermediate values between the values of the plant with intermediately and least growth. The highest spore count was observed in the plants inoculated with Glomus Jasciculatum 20g + single super phosphate 109 followed by Glomus Jasciculatum20 g + Azospirillum 20 g + single super phosphate 10 g. With regard to the root infection, plants inoculated with Glomus Jasciculatum 5 g + Azospirillum 20 g + single super phosphate 109. and Glomus Jasciculatum 20 g + Azospirillum 109 + single super phosphate 109 revealed maximum percentage of infection. The Azospirillum population was maximum in the plants inoculated Glomus fasciculatum 5 g + Azospirillum 109 + followed by Glomus mosseae 20 g + Azospirillum 20 g + single super phosphate 109. The standard procedure for identification and quantification of abscisicacid (Little et al., 1972) was modified, as clear banding patterns were not obtained. Using the modified procedure, the characteristic-banding pattern corresponding to standard abscisic acid was obtained and confirmed when standards of abscisic acid were also simultaneously run with samples. Banding patterns and quantification of samples in each of the four age groups, the growth regulator treatments and the treatment with arbuscular mycorrhizal fungi and Azospirillum inoculated plants were also successfully conducted.

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