Cytomorphological studies and development of protocol for tissue culture in Black Caraway (Bunium persicum Boiss)
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Date
2008
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SKUAST-K
Abstract
Six ecotypes of Black Caraway (Bunium persicum Boiss) adapted to different agro climatic conditions of the state were selected for cytomorphological studies and development of protocol for tissue culture. Analysis of variance revealed significant to highly significant differences among the ecotypes for different traits. Both phenotypic and genotypic coefficient of variation were moderate for number of seeds tertiary umbel-1, seed yield plant-1 and 1000-seed weight. It was low for all other characters. Heritability in broad sense was high for the characters like number of secondary branches plant-1, days to initial, 50 per cent flowering and full bloom, number of umbels plant-1, number of umblets primary-1, secondary-1 and tertiary-1 umbel, umbel diameter of primary and secondary umbels, days to 80 per cent physiological maturity, number of seeds primary-1, secondary-1and tertiary-1umbel, seed yield plant-1 and 1000-seed weight. Correlation of important characters like seed yield plant-1 was significant and positive with plant height, number of primary branches plant-1, umbel diameter of primary and secondary umbels, number of umbels plant-1, number of seeds primary-1 and secondary-1 umbel and 1000-seed weight, but significant negative correlation with number of secondary and tertiary branches plant-1. 1000-seed weight has significant and positive correlation with plant height, number of primary branches plant-1, 80 per cent physiological maturity and number of seeds primary-1 umbel and negative correlation with number of secondary and tertiary branches plant-1 , number of umbels plant-1, number of seeds secondary-1 and tertiary-1 umbel. The seeds of Chrar-e-Sharief and Vasturvan ecotypes were bolder than the seeds of Gurez and Drass ecotypes. The seed size of Harwan and Paddar ecotypes was intermediate in size. Seeds of Chrar-e-Sharief, Vastuvan, Harwan and Paddar ecotypes were longer (more selendrical) than Gurez and Drass ecotypes. Essential oil content was higher in Chrar-e-Sharief ecotype whileas it was lower in Drass and Gurez ecotypes. The oil content of Vasturvan, Harwan and Paddar was lower than Chrar-e-Sharief ecotype but higher than Drass and Gurez ecotypes. Seeds harvested at 80 per cent maturity had higher essential oil content as compared to seeds harvested at full maturity. It was also observed that although Gurez and Drass ecotypes had low quantity of essential oil as compared to Chrar-e-Sharief ecotype but they were richer in important oil constituents with respect to flavour. Disease incidence of leaf blight, umbel blight and tuber rot was recorded to be higher in Chrar-e-Sharief ecotype, whileas it was recorded minimum on Gurez and Drass ecotypes. It was also observed that disease incidence progressively increased from vegetative to seed maturity stage. Cytological studies revealed that Black Caraway (Bunium persicum Boiss.) is a diploid species with 2n=14 chromosome number. In the protocol development for tissue culture in Black Caraway (Bunium persicum Boiss), it was observed that nodal segments are the best explant source for tissue culture. The best sterilant for sterilization of explant proved to be 20 per cent sodium hypochlorite for 15 minutes. The explants were cultured on 4 media types. It was observed that MSN medium containing 0.4 mg l-1 2, 4-D was the best for callus initiation and establishment. The callus proliferation was maximum on MSN medium containing 0.4 mg l-1 2,4 D and 2.25 mg l-1 kinetin. It was evident that callus growth had an upward trend up to 4 weeks of incubation after which the growth of callus was slow. Shoot induction was best when MSN medium was supplemented with 2.25 mg l-1 kinetin or 3.00 mg l-1 BAP and without auxin. Maximum number of microshoots developed from green callus morphotype. Maximum rooting of microshoots was possible when half strength MSN medium containing 6 percent sucrose was supplemented with 0.1 mg l-1 NAA. Maximum micro tuber formation was achieved on half strength MSN medium containing 1.00 mg l-1 IBA or 0.20 mg l-1 NAA.Maximum induction of somatic embryos was achieved on MSN medium containing 0.05 percent activated charcoal. It was observed that maximum number of somatic embryos developed from yellow callus morphotype. Maturation of somatic embryos was achieved when MSN medium was supplemented with 0.05 mg l-1 ABA and 1.00 mg l-1 AS. Chilling and short photoperiod regime was found to be necessary for the germination of somatic embryos. Chilling of embryogenic callus (4±1oC) and short photoperiod regime (8:16h) was found suitable for germination of somatic embryos. The mature somatic embryos germinated on MSN medium containing 1.00 mg l-1 GA3 and IBA plus AS (0.50 mg l-1 and 5.00 mg l-1 , respectively). Successful hardening of microplants was achieved in a mixture of soil, sand and FYM (1/3:1/3:1/3) under regime- I nutrition.
Description
M.Sc Thesis submitted to SKUAST Kashmir
Keywords
Bunium persicum Boiss, callus, ecotyp, somatic embryo, Genetic and Plant Breeding