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ThesisItem Open Access Refinement of in vitro propagation technique in pineapple var. mauritius and mass multiplication of elite clones(Department of pomology and floriculture, College of horticulture,Vellanikkara, 1996) Jo Jose C; Radha TThe studies on refinement of in vitro propagation technique in pineapple var. Mauritius and mass multiplication of elite clones were conducted at Kerala Horticulture Development Programme, Kerala Agricultural University, Vellanikkara during 1993 – ’95. Based on the survey conducted at the major pineapple growing areas, namely, Vazhakulam – Muvattupuzha regions of Ernakulam district, five elite clones of Mauritius variety with higher yield and other desirable fruit characters were selected propagules from the selected clones were planted at Vellanikkara for taking explants for their mass multiplication through refined in vitro propagation techniques. For shoot tip explants, treatment with emisan 0.1 per cent for 35 minutes followed with mercuric chloride 0.1 per cent for 10 minutes and for lateral bud explants treatment with emisan 0.1 per cent for 10 minutes followed by 0.1 per cent mercuric chloride for three minutes was found to be the ideal surface sterilization treatment. Culture establishment and growth initiation of shoot tip explants from different sources (suckers, crowns and slips) and lateral bud explants were better in MS medium supplemented with BAP 3 and 4 mg 1-1. Explants from shoot tips were found to be better than those from lateral buds for achieving faster culture establishment and growth initiation. Enhanced release of axillary buds was the maximum in Ms medium containing BAP 4.0 mg 1-1. Among the different subculture stages, multiple axillary bud production was higher in second suculture stage. Addition of casein hydrolysate 100.0 mg 1-1 in to the medium with BAP 4.0 mg 1-1 favoured the production of axillary buds. Adventitious bud initiation from sucker shoot tips was fastest in MS medium supplemented with BAP 5.0 mg 1-1 + NAA 1.0 or 0.5 mg 1-1. For adventitious bud production from lateral buds, treatment with BAP 7.5 mg 1-1 + NAA 1.0 mg 1-1 was the best. Proliferation rate of adventitious buds was maximum in MS medium supplemented with BAP 4.0 mg 1-1 + NAA 0.5 or 1.0 mg 1-1. Rate of multiplication of adventitious buds was higher in liquid medium under shake culture condition, than in solid medium. Faster shoot regeneration and increased vigour of the shoots were resulted in growth regulator free MS medium. However, highest number of shoots were produced in MS medium supplemented with BAP 1.0 mg 1-1. Liquid medium under shake culture condition was found superior of solid media with respect to initiation of shoots and mean number of shoots, however, the latter resulted in longer shoots. In vitro rooting was fastest in MS medium, without any growth regulator, which produced longer and normal roots with secondaries and root hairs. Though addition of NAA (3 mg 1-1) resulted in increased number of roots, they were very short and hair like. Stationary liquid medium was found superior to solidThesisItem Open Access Floral biology and compatibility in gladiolus(Department Of Pomology And Floriculture,College Of Horticulture,Vellanikkara, 1999) Jisha, V; KAU; Suma, AThe present investigation on 'Floral biology and compatibility in gladiolus was conducted in the Department of Pomology and Floriculture, College of Horticulture, Vellanikkara during the period from July 1997 to December 1998. The investigations on floral biology, pollen studies and the extent of compatibility between different varieties were done in the experimental field of AICRP on Floriculture. All the varieties showed significant variation among themselves for all the characters studied. Accession-l showed maximum corm germination percentage (94.89), plant height (68.86 cm), leaf number (9.44), spike length (64.67 cm), spike yield per plant (3.11) and took minimum days for spike emergence (46.56) and first flowering (60.44 days). These desirable qualities combined in a single variety indicate its scope for using in further breeding programme. American Beauty had maximum duration of flowering (12.44 days) and highest cormel number (8.11). Agnirekha had maximum number of florets per spike (15.00) and corm weight (31.66 g). Mansoer Red had highest spike weight (25.63 g) and Wedding Bouquet had maximum floret size (11.14 cm) and cormel size (1.84). Maximum number of florets per spike was observed in Pacific. It had longest vase life (8.66 days), corm size (5.18 cm) and conn number (1.78) also. Tambri showed maximum cormel weight (4.65 g). In all the varieties fullanthesis occurred from 7.15 a.m. and peak time was 9.00 a.m. Anthers dehisced within two to three hours after unfurling of perianth .. Stigma became receptive only after it turned feathery. By 9.30 a.m. on the day of anthesis stigma turned feathery and after that upto two days it remained receptive. White Friendship had maximum pollen fertility (85.90 %) and pollen production per anther (39500) while Pacific registered highest pollen diameter (38.09 J.!). Maximum pollen germination was observed in 15 per cent sucrose + 75 ppm boric acid medium for the two varieties studied viz., White Friendship and Accession-L Pollen viabil ity was the highest when stored in desiccator at 4"C. High heritability and high genetic advance were observed in characters I ike days to first flowering, spike length and leaf area: This shows that most likely the heritability is due to additive gene effects and selection may be effective for these characters. PCV and GCV were high for leaf area, spike yield per plant, number of florets open at a time and duration of flowering and they were low in the case of vase life, spike length, plant height and leaf number. The number of florets per spike which determines the market quality had positive correlation with spike weight, number of florets open at a time and leaf area. So the number of florets per spike can be increased with the improvement of any of these characters. A straight away selection from - gennplasm lines will be very effective for making improvement in this character. The genotypic correlation coefficients were found to be higher than the phenotypic correlation coefficients for most of the characters. This indicates that the environment had lesser effects on these characters. The direct and indirect effects of 12 characters on number of florets per spike showed that spike weight had the maximum direct effect on number of florets per spike: It was followed by duration of flowering, collar girth, spike length and vase life. The direct selection for the characte-s like spike weight and duration of flowering would be beneficial for crop improvement, since these two also showed positive correlation coefficients. Negative direct effects on number of florets per spike was from days to first flowering, plant height, floret size, leaf area, leaf number and spike yield per plant. The highest positive indirect effect on number of florets per spike was of leaf area through spike weight. The indirect effect of duration of flowering on number of florets per spike through number of florets open at a time was the lowest. Ten gladiolus varieties namely, White Friendship, Tambri, Echo Saunder, American Beauty, Tiger Flame, Wedding Bouquet, Accession-l , Accession-2, Amal and Pacific were selected and crossed in all possible combinations. In Amal and Tambri there was no pod development and seed set in both cross pollination and self pollination tests. In self pollination, pod set, number of seeds per pod and weight of seeds per pod were higher than that in cross pollination. Pods took 25 to 35 days to mature. In self compatibility Accession-I had maximum -pod set (100 %) and highest number of seeds per pod (77.21). Weight of seeds . per pod was highest in Wedding Bouquet (1.36 g). In cross compatibility, the pod set was maximum in Accession-2 x White Friendship (91.78 %) and the number of seeds per pod was highest in Accession-? x Accession-I. Thc weight of seeds per pod was maximum in Tiger Flame x Amal (0.92 g). The seed germination percentage was maximum in Pacific (78.94 %) in self compatibility studies. In cross compatibility the seed germination percentage was highest in Accession-2 x White Friendship. The varieties with highest number of florets per spike, vase life and early flowering were observed in this investigation. Also the compatible varieties were identified. These varieties can be exploited in further breeding programmes in gladiolus.ThesisItem Open Access Vegetative, Floral and fruit characters in mangosteen (Garcinia mangostana L.)(Department of Pomology and Floriculture, College of Horticulture, Vellanikkara, 1996) Ajay Alex; KAU; Sarah, T GeorgeThe present investigations on the growth habit, phases of growth, flowering and floral biology, fruit development, seed viability and storage life of mangosteen were carried out in the Department of Pomology and Floriculture, College of Horticulture, during the period 1994-96. The studies indicated that shoot growth in mangosteen coincided with the main flushing season from June to August and with a second one from January to February. Maximum shoot growth was observed during July. The growth of the tree was slow, with an extension growth of 6.91 cm in an year. The tree had a monopodial orthotropic trunk meristem which showed continuous growth. Laterals exhibited plagiotropy, sympodial growth and sylleptic branching habit. Leaf arrangement was spiral in the seedling stage but distichous on the branches of mature tree. Emerging leaves which were purplish red, later changed to dark green. The flowering season was from December to January. But development was completed in 28 days. Flowers were female and borne terminally on branchlets either singly or in groups of two to four. Flower drop was meager. Peak anthesis period was between 17.30 and 18.00 hours. Flowers had four scarlet red sepals and four yellow petals each having imbricate aestivation. Androecium consisted of 18-20 staminodes. Gynoecium was syncarpous with five to seven carpels having single ovule in each locule on axile placentation. Style was short and had a five to seven fid capitate stigma at its end. Anthers failed to dehisce until flower opening but a few showed signs of dehiscence after anthesis. Stigma showed no signs of receptivity. Anthers produced numerous non viable pollen grains which failed to germinate in vitro. Different methods of pollination had no effect on fruit set. Initial set was high but a fruit drop of 41 per cent occurred during first month. Though fruit development was parthenocarpic and seed development parthenogenetic, seeds produced were viable. Therefore, mangosteen can be considered as an obligate agamosperm where proembryos developing from integuments of embryosac mature into embryos. Pulp development took place from 42nd day onwards. Average weight of ripened fruit was 100 g. The percentage contribution of pulp towards total fruit weight at ripening stage was 33.00 per cent as against 62.30 and 4.70 per cent in the case of rind and seed, respectively. Chemical composition of pulp showed a decreasing trend. Total sugars, reducing sugars, non reducing sugars and sugar : acid ratio increased upto harvest. Season of harvest coincided with South West monsoon. Stage of harvest was identified as 90 days after fruitset. Such fruits ripened normally in two days at ambient temperature and showed no difference in quality as compared to that of tree ripened fruit. At this stage, 25 per cent of the fruit skin developed a purple colour and scar formed at the stalk end was smooth, without any exudation of gum. Mechanical injury should be avoided during harvesting and handling to save fruits from Transluscent Flesh Disorder. Yield varied from 650 to 3350 fruits/tree. Number of segments, which was same as that of stigmatic lobes, ranged from four to seven. However, number of viable seeds ranged from zero to three. Fruits caught in the rain were severely affected with gamboges, a disorder, which accounted to about 33.82 per cent fruit loss. Exudation of yellow gum from the rind was the characteristic symptom. The fruit pulp also became yellow, gummy, corky, bitter in taste and inedible. Biochemical analysis showed that ripened fruit contained water 76.57, protein 0.5, citric acid 0.32, total sugars 17.02, reducing sugars 3.22, non reducing sugars 13.80, nitrogen 0.28, phosphorus 0.01, potassium 0.13, calcium 0.01 and magnesium 0.24 on percentage basis. Sugar : acid ratio, TSS and ascorbic acid content was 53.18, 27.00 0brix and 5 mg/100 g, respectively. β carotene was only in traces. Fruits stored under refrigerated conditions showed no quality deterioration and fruit loss even after one month of storage. Fruits kept in bamboo baskets lasted for a fortnight. Keeping quality of fruits even without any treatment was more than a week. During storage TSS, sugars and sugar: acid ratio decreased, whereas acidity increased with the storage period. Seeds varied in size and shape. Viability was very high when sown immediately after harvest. Storage reduced the viability and was completely lost by 35 days of storage. Seeds took 20 days for germination. Germination was hypogeal with single seedlings arising normally, but 10 per cent polyembryony with 2-4 seedlings/seed was also noticed.ThesisItem Open Access Micropropagation of phalaenopsis(Department of Pomology and Floriculture, College of Horticulture, Vellanikkara, 1996) Jyothi, Bhaskar; KAU; Rajeevan, P KInvestigations were carried out at the Plant Tissue Culture Laboratory attached to the All India Co-ordinated Floriculture Improvement Project (AICFIP), College of Horticulture, Vellanikkara, during 1993-96 to standardise the micropropagation technique in Phalaenopsis. Out of the different explants tried, response was shown by inflorescene stalk node, inflorescence stalk tip and pollinia collected from the field grown plants and apical bud, shoot node, basal portion, leaf and root of plantlets grown in vitro. Maximum survival (40%) of nodal explants was recorded at the sterilant combination involving mercuric chloride (0.01%) for 30 min., streptomycin + pencillin (0.01%) for 90min., followed by final sterilization using mercuric chloride (0.10%) for 10 min. For flower bud, the combination involving emisan (1.00%) for 30 min. followed by alcohol (50%) for 1 min. recorded the maximum survival percentage (55). The ½ MS liquid medium containing BA 5 ppm+ NAA 2ppm + 2, 4-D ppm + CW 15 percent recorded the minimum number of days for nodal swelling and bud development (6 and 14 days, respectively). Basal portion was found to be the best explants with respect to rate of increase in shoot number (8.00) and leaf number (7.67), followed by shoot node. The latter recorded the highest root number (7.67) at the end of 12 weeks of culturing. As to the position of explant, highest increase in the length of the buds was recorded for first node, followed by second, third and fourth. Full strength MS and KC media were far inferior to ¼ MS, ½ MS, ¾ MS and VW media, for culturing. Maximum number of shoots (5.00) and leaves (7.67) were recorded for ¼ ms AND ½ ms media after 8 weeks of culturing. Physical state of the medium, viz. liquid and semi –solid did not show any significant difference. Sucrose at 1.5 percent level recorded the maximum number of shoots and leaves after 8 weeks of culturing. Thiamine – HCL increased the shoot and leaf number at 20ppm level at the end of 8 weeks, whereas the presence did not favour the production of roots. The time taken for callusing in pollinia was minimum (2.0 days) at BA 5 ppm + NAA 2 ppm + 2, 4-D 2ppm in ½ MS medium containing 3 percent sucrose. When 90 day old pod was used, protocorm formation was observed in ½ MS medium containing BA 10ppm + NAA 1 ppm and KIN 5 ppm +2, 4-D 2ppm. When the effect of NAA was considered root production was increased at the combination NAA 5 ppm + BA 10ppm + adenine 10 ppm. With regard to the effect of BA, that at 25ppm in combination with adenine 10 ppm + NAA 1 ppm recorded maximum number of shoots and leaves. Shoot and leaf number was maximum at ¼ MS medium containing BA 20 ppm+ 2, 4-D 2.5ppm, whereas root production was maximum at BA 20ppm + 2, 4-D 5 ppm. As to the combined effect of BA, NAA and 2,4-D in ¼ MS medium, the combination BA 5ppm + NAA 2 ppm + 2,4-D 2 ppm recorded the maximum number of shoots and leaves. Different levels of KIN, was found to have no significant influence on the production of shoots, leaves and roots. When the combined effect f KIN, NAA and 2, 4-D was considered ¼ MS medium containing KIN 20ppm + NAA 2 ppm + 2, 4-D 2 ppm recorded maximum number of shoots and leaves. The apical bud did not show any multiple shoot production during the 12 weeks culture period as influenced by coconut water, whereas first node at CW 10 percent, shoot node at CW 10 per cent, shoot node at CW 15 percent, and basal portion at CW 25 percent recorded the highest number of shoots.Maximum number of leaves was produced by apical bud at CW 20 percent , first node and shoot node at CW 10 percent, and basal portion at CW 25 percent. Both tender and mature coconut water were equally effective. Fresh and upto 6 days old coconut water could also be used with similar effect. Peptone at 1000 ppm was found to influence favourably the induction of multiple shoots from in vitro shoots. Culturing the nodal explants in liquid media with filter paper bridge or keeping in the dark were found to reduce phenolic blackening. With the increase in the concentration of antioxidants, there was a proportionate reduction in media discolouration. Activated charcoal and triadimefon added in the media were found to influence the root production from shoots. Length of the root was maximum at triadimefon 20 ppm. Sucrose at 1.5 percent level recorded the minimum number of days for PLB development and the maximum number of PLB’s developed. Thiamine – HCL, coconut water, tomato juice and peptone did not significantly influence the time taken for PLB formation, but favoured the number of PLB’s developed. With regard to PLB formation from shoot node, ½ MS medium containing BA 5 ppm + NAA 2 ppm + 2, 4-D 2 ppm recorded the minimum number of days for PLB development and maximum number of PLB’s. When PLB formation from in vitro leaf was considered, cent percent of the leaf cultures developed PLB’s at the combination BA 25 ppm + adenine 10 ppm + NAA 1 ppm and the time taken for PLB formation was minimum. Cent percent of the cultures developed PLB’s from in vitro roots at BA 25 ppm + adenine 10 ppm + NAA 1 ppm and the time taken for PLB formation was minimum. Further growth of PLB’s and Plantlet development was the best in ¼ MS medium containing BA 15 ppm +NAA 1 ppm followed by adenine 8 ppm + BA 16 ppm. Regarding the effect of coconut water, PLB growth at CW 15 percent and plantlet development at CW 25 percent recorded the best results. Light favoured plantlet development, multiple shoot formation and PLB formation from shoot node and in vitro root, whereas dark period favoured early development of PLB’s from in vitro root, whereas dark period favoured early development of PLB’s from in vitro leaf, callusing and PLB proliferation. Healthy, large and robust plants were produced when plantlets were grown in 250 ml conical flask, followed by large test tubes. When the plantlets grown previously in medium triadimefon 20 ppm were hardened by spreading over sterile charcoal pieces for two weeks, planted in coconut husk and were hung in the orchidarium with high humidity, cent percent survival was recorded even after 8 weeks of planting out. The nutrient solution 30:10:10(0.50%) and 17:17:17 (0.10%) recorded the highest survival percentage after 12 weeks of planting out. The growth characters of the plants. Viz., plant height, leaf number, leaf length and width, root number and root length were found to be maximum for the plants sprayed with 17:17:17 at 0.10 percent level. The survival percentage of plants showed a slight decrease with time and all the plant characters increased with time except the number of leaves.
