Development of High Yielding Saffron Mutant I.A. Khan G.B. Pant University of Agriculture & Technology Saffron Cultivation and Marketing Project Chaubatia-263651 Almora India Keywords: 5-Branched stigma, mutation, triploidy Abstract Saffron (Crocus sativus L.) is a temperate, medicinal and low growing herb. Its dried stigma is used as Bio-medicine, dyes and perfumes. All allies of genus Crocus are diploid but Crocus sativus L. is a triploid in genetic make up (2n = 3x = 24). Due to triploidy, meiosis in C. sativus is highly erratic and genetically unbalanced gametes are formed which lead to the formation of sterile gametes and ultimately no sexuality is involved which is an essential phenomenon for seed production. Due to absence of sexuality in Saffron, a non conventional mutation breeding was initiated to develop improved mutants of Saffron at Horticulture Research Centre, Chaubatia, Almora, India, located in Himalayan region. Corms with different doses of gamma rays (Co 60 ) were subjected to induce the variability in Saffron. Five sets, each consisting of 100 uniform corms of 5-6 g in weight, were irradiated with 0.5, 1.0, 1.5, 2.0 and 2.5 doses of gamma rays. Variability in respect of sprouting time, plant height, induction of flowering, shape of petals and number of filaments in a stigma was noted. Plants exhibiting mutations were identified and bulbs produced from mutants were harvested separately to further the generations. An outstanding mutant which was isolated was, a mutant with five branched stigma, instead of three as in normal plants. On evaluating this mutant, it was found superior in respect of number of branches in stigma. Average length of filaments in five branched stigma noted to be 2.75 cm whereas in three branched stigma it is 2.70 cm. Dry weight of normal three branched stigma is 5 mg. whereas in five branched stigma it was noted to be 6.950 mg which is higher than control. Development of 5 branched stigmas is result of gene mutation due to induction effect. INTRODUCTION Saffron (Crocus sativus L.) is a temperate medicinal and low growing herb. Its dried stigma is used in Bio-medicine, dyes and perfumes. More recently Saffron s use as antitumour, cardiovascular protective agent and use of extract of Saffron corms against malignant cells indicate the importance of this plant (Escribano et al, 1999; Abdullaev, 2002). Saffron plant is sterile and has no capability of proudcing seeds. Propagation is made by under ground vegetative part called corm. Genetically it is triploid in make up (2N=3x=24). Due to triploidy, no recombination of genes occurs and consequently no variants are developed. Moreover transportation, storage and packing of corms increase the cost of cultivation. Further more, corms are also affected by virus which deteriorate quality of Saffron. Keeping importance of the plant and affecting factors for cultivation in view, present investigation was undertaken to induce the variability
by physical irradiation. Main objectives of employing physical irradiation were development of polyploid forms and tetrafid or pentafid or hexafid stigmatic plants. MATERIALS AND METHODS Experiment was conducted during 2004-2005 at Chaubatia garden, Horticulture Research and Extension Centre, Chaubatia, located at 6000 feet height from sea level in Central Himalaya. Five sets, each consisting of 100 uniform corms and each corm 5-6 gm in weight, were irradiated with 0.5, 1.0, 1.5, 2.0 and 2.5 doses of gamma rays at N.B.R.I., Lucknow. A set of 100 untreated corms was used as control. After irradiation, corms were planted in raised beds at 10x15 cm dimension and at 8-10 cm deep. Beds were covered with grass mulch. All cultural practices were made to raise the normal crop. Observations on sprouting, plant height, days to flower, flower size, flower shape, flower colour, flowering percentage, branches of stigma and weight of dry stigma were taken into consideration. Individually all plants of VM1 generation were examined and identified mutants were harvested to advance the next generation. RESULTS AND DISCUSSION Effect of irradiation was studied on the basis of various morphological characters. Induction of early sprouting in 0.5, 1.0 and 1.5 and delayed sprouting in higher doses was observed. Induction of flowering and plant height in lower doses and decrease in plant height in higher doses was also observed. Higher percentage of sprouting in lower doses and poor sprouting in higher doses was also noted. Cent percent survival in 0.5, 1.0 and 1.5 and reduced survival percentage in higher doses was also recorded (Table 1). On the examination of treated population, serrated petal mutant plant and 3-petal plant was identified and a plant having five branched stigma, instead of three as a normal plant was found. Average length of filament in five branched stigma noted to be 2.75 cm whereas in three branched stigma it is 2.70 cm. Dry weight of normal three branched stigma is 5 mg whereas in five branched stigma it was noted to be 6.950 mg which is higher than normal (Table 2). Corms of identified mutants (petal mutants and five branched stigma mutant) were harvested separately and advanced to VM2 generation to bring them true to type form and also to compare, with parental lines. All identified mutants including mutation in petals and stigma number failed to inherit the characters indicating reversible mutation. No polyploid forms were observed. Gamma irradiation affects the sprouting, growth and flowering of the plants. Delayed sprouting, slow growth in higher doses, increase/decrease of plant height at lower and higher doses respectively and induction of flowering in VM1 generation at lower doses are due to auxin synthesis whereas at higher doses it is inhibited resulting into delayed and stunted growth. Present findings are in conformity with the reports of others (Gordon, 1957; Khan, 2003). Stunted growth, reduction in survival and reduced fertility is also attributed to genetic loss due to chromosomal aberrations and gene mutation (Sparrow et al, 1961; Datta and Gupta, 1980; Khan, 2003). Flowering induction in lower doses of gamma irradiation in VM1 generation is result of mutation in bio-chemical pathway which assists in the synthesis of flower inducing substance leading to formation of flower whereas same process is not inherited in the next generation. Formation of five branched stigma is result of somatic mutation where apical cell responsible for making three branched stigma divide in five cells instead of three,
due to induction effect. Plant with five branched stigma also reported to be found some time in natural growing conditions which is also result of environmental somatic mutation. In present investigation we failed to produce permanent mutation but mutant with heritable five branched stigma if developed, definitely increase the yield of Saffron. Literature Cited Abdullaev, FI. 2002 Cancer chemopreventive and tumoricidal properties of Saffron (Crucus sativus L.) Exp. Biol. Med. (May wood) 227(1):20-25. Datta, S.K. and Gupta, M.M. 1980. Effect of gamma irradiation on rooted cuttings of small flowered chrysanthemum. New Botanist VII:73-85. Escribano, J., Rios, I. and Fernandez J.A. 1999. Isolation and cytotoxic properties of novel glycoconjugage from corms of Saffron plant (Crocus sativus L.). Biochim Biophys Acta 1426(1):217-222. Gordon, S.A. 1957. The effect of ionising radiation on plants: Biochemical and physiological aspects. Quart Rev. Biol. 32:3-14. Khan, I.A. 2003. Induced mutagenic variability in Saffron (Crocus sativus L.) Proc. 1 st International Symposium on Saffron Biology and Biotechnology, Albacete, Spain. Oct 22-25, 2003 P. 21. Sparrow, A.H., Gunay, R.L., Micksche, J.P. and Schairer, L.A. 1961 Some factors affecting the responses of plant to acute and chronic radiation exposure. Rad. Bot. 1:10-34.
Table 1. Effect of Irradiation on various characters. S. No Characteristics Control 0.5 1.0 1.5 2.0 2.5. 1 Days to sprouting 42 39 40 41 50 55. 2 Days to flowering 65 60 64 66 - -. 3 ( Plant height (cm 12.25 13.50 14.50 13 11 10. 4 (%) Sprouted bulbs 100 100 100 100 75 60. 5 (%) Survival plant 100 100 100 100 60 49. 6 Flowering plants (%) 40 50 60 55 - - Table 2. Data on 5 branched stigma mutant. S. No Characteristics Control Mutant. 1 ( Average length of stigma (cm 2.70 2.75. 2 ( Weight of dry stigma (mg 5.00 6.950