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Birsa Agricultural University, Ranchi
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ThesisItem Open Access Protocol Development of In- Vitro Cultivation of Bamboo ( Dendrocolamus As per )(Birsa Agricultural University, Ranchi, 2002) Peddy Srikanth; Z.A. HaitherBamboos are perennial, woody and evergreen monocotyledonous arborescent grasses belonging to the family Poaceae (Graminae) and Sub-family Bambusoideae. There are about 75 genera and 1250 species of bamboos. Dendrocalamus asper is one of the economically important and widely cultivated bamboo species. The tender shoots of this plant can be consumed as food and is a good source of foreign exchange to various countries. Mature culms of this plant are used for pulp and paper manufacture. Bamboos are propagated artificially by different methods, including through seed. But utilization of seeds as propagating material is difficult and unreliable due to long and unpredictable flowering habit, short dormancy period of seed, poor viability, inborn microbial infestation, poor seed set during off-season flowering, seed sterility and large scale. Consumption of seeds by rodents and wild animals. The vegetative methods, on the other hand, are costly, lobour intensive, cumbersome and time. Consuming. These vegetative propagates are bulky, difficult to transport to distant places and their survival rates are also not very high. This limits large scale cultivation of bamboos in general. Under the situation, propagation through tissue culture seem to be a viable method for large scale propagation of the bamboo species. Therefore the present project was undertaken to establish a protocol for in-vitro propagation of Dendrocalamus asper. In the present study nodal segments (3-4cm) with axillary buds from young juvenile mother plant was used as explants. Surface sterilization using 0.1% (w/v) mercuric chloride (Hgcl₂) for 10 minutes followed by 3-4 times subsequent washing with sterile distilled water proved the best as it resulted the highest percentage (92.68%) of bud break after two weeks. The sterilized nodal segments were cultured aseptically on MS medium supplemented with 0-15 mgl-1 BAP and maximum shoot proliferation. (14-15 shoots per propgule) was achieved on medium supplemented with 12mgl¹ BAP. These proliferated axillary shoots were excised and subcultured on MS liquid medium +3 mgl BAP for the first two subcultures to increase the number of shoots. The shoot multiplication was achieved on both MS solid as well as liquid medium supplemented with 1-5 mgl¹! BAP Highest rate of shoot multiplication (fold) i.e., 15.77 was obtained on MS liquid medium supplemented with 3 mg l-¹ BAP in four weeks. MS solid medium supplemented with 3 mgl¹ BAP resulted only 8.55 fold. Incorporation of NAA (0.2-1.0 mgl ¹) to the medium along with BAP did not increase the rate of shoot multiplication and shoot length but it resulted in better quality erect shoots. MS medium in its full strength (1x) was found to be the most effective basal nutrient medium for shoot multiplication. The studies on sucrose concentration in the medium showed that 3% sucrose was essential for rapid multiplication of shoots. The effect of pH reflected that shoot multiplication occured even on acidic medium and highest rate of shoot multiplication (15.88) was obtained at pH 5.8. A regular subculture cycle at an interval of 4 weeks resulted in healthy cultures devoid of brown leaves and high rate of shoot multiplication. For in-vitro root regeneration on MS medium supplemented with 10mgl-¹ IBA yielded 90% rooting, 19.66 roots per propagule in four weeks, while 3 mg NAA supplemented MS medium resulted 91.66% rooting with 10 roots per propagule NAA resulted short roots while IBA resulted long roots. Addition of BAP (0.1-0.5 mgl ¹) to the rooting medium, neither enhanced root regeneration percentage nor improved the number of the in-vitro roots The cultured plantlets were successfully hardened under high humidity on sterilized soil sand FYM(1:1:1) mixtured with 1/2 strength MS nutrient medium irrigations (without organics).ThesisItem Open Access Protocol Development of invitro Clonal Propagation of Orchid ( Vanda Spa)(Birsa Agricultural University, Ranchi, 2002) Ranjeet Kumar Sinha; Z.A HaitherOrchids, one of the most beautiful group of flowering plants belong to the family Orchidaceae (Monocotyledons). The exquisite beauty of Orchid flowers due to brilliance in colour, remarkable range of sizes, manifold shapes, and variation in the form and wide range of distribution has aroused highest admiration throughout the world. The Orchid comprises about 800 genera with around 35,000 species. In India, about 1300 species of Orchids are found in Himalayas and others scattered in eastern and western Ghats. A vast majority of Indian Orchids are confined to mountain where they are distributed from base of hill to the elevation of 4300 m above mean sea level in climates ranging from tropical to temperate. Orchids are terrestrial, epiphytic, lithophytic or saprophytic but no Orchid is aquatic. The cut flower trade of Orchid involves 3% internationally. Major suppliers, like, Thailand, Netherlands and Singapore export flowers worth of US $ 80.0, 77.4 and 20.0 millions, in order per year. Due to their alkaloid, flavanoid, glycosides and other phytochemical constituents Orchids have high therapeutic value. The flower juice of Vanda coerulea is used to cure eye diseases. Cymbidium elegans, Cymbidium pubescens, Epicactic latifolia are used as local medicines. for treatment of nervous disorders. Orchids are also used in many countries as food or for making refreshing drinks. Unfortunately the natural population of Orchid is fast declining due to excessive collection and over harvesting by traders and botanical explorers. So there is need to cultivate and conserve the endangered Orchids. The conventional method of propagation is tedious and time taking. The alternative means of propagation is in vitro clonal propagation. Keeping this in mind the present experiment. on developing a viable protocol for in-vitro clonal propagation of Orchid (Vanda Miss Joaquim) was undertaken. The explant, like, shoot apex and shoot node were washed with detergent and teepol and then sterilized with 0.2% mercurio chloride for 10 minutes. The explants were cut in small pieces under laminar flow hood and subsequently inoculated in Murashige and Skoog (1962) medium modified with different plant growth regulators. The inoculated materials were cultured under aseptic condition at 25+2°C with 16 hours photoperiod of 3000 lux. The medium containing 2% sucrose, 2 mgl¹ BA+ 0.2 mgl¹ NAA was best for shoot node culture and developed 10 shoots/node and 4 leaves per shoot. Protocorm like bodies were developed in cytokinins combination. The combination 1 mgl BA + 0.3 mgl kinetin proved better for getting higher number of buds. However, 7.67 buds/node were found with 1mgl kinetin + 0.1 mgl¹ 2,4-D in around 46 days. It is worthy to note here that 2 mg1¹ kinetin in absence of 2,4-D yielded 7.56 buds/node which is statistically at par with the combination treatment 1mgl kinetin + 0.1 mgl 2,4-D. The shoot apex culture gave significant results on MS. medium supplemented with 2 mgl¹ BA + 0.5 mgl'¹ NAA, 1 mgl¹ BA + 0.2mgl kinetin and 1mgl kinetin + 0.1 mgl 2,4-D. Sub-culturing of plantlet on 2 mgl¹ BA and 0.5 mgl¹ NAA gave about 70-100 shoots. Best result on rooting was achieved on MS medium supplemented with 1 mgr¹ IBA+ 0.5 mgl¹ NAA, 1 mgr¹ NAA + 0.1mgl¹ BA. The maximum root length (49.5 mm and 60 mm) was obtained on medium supplemented with 1 mgr¹ NAA+ 0.1mgr¹ BA and 2 mgl¹¹ NAA + 1 mg/¹¹ IBA respectively. The cultured shoots were hardened successfully in pots containing bark, brick pieces and charcoal in 1:1:1 ratio.ThesisItem Open Access Protein Profiling of Rice (Oryza sativa L.) Under Micronutrients Stress Condition(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2007) JHA, APARNA; Haider, Z.A.Today in most countries it is chemical fertilizers which are supplying most of the macronutrients needed by crops. Farmers seldom apply micronutrients, even though intensive modern agriculture has a depleting effect. Yields are higher than those of the past, while early maturing varieties make it possible for farmers to increase the number of crops grown in the course of a year. Micronutrients are being continuously removed in high yields of harvested produce, without being replaced. The result is widespread micronutrient deficiencies. Deficiency is shown in various kinds of physiological damage, all of which affect the quality and quantity of produce. Molybdenum deficiency often results in whiptail of cauliflower, yellow-spot of citrus and blue-chaff of oats; examples of boron deficiency are hollow heart of cauliflower, stone fruit of papaya; zinc deficiency causes khaira disease of rice etc. The present study dealt with identification of proteins which are specifically altered (induced or suppressed) in response to micronutrients stress (deficiency and toxicity) as well as Aluminium stress (toxicity). One dimensional gel electrophoresis (followed by silver staining) was used to identify proteins altered in rice tissue in response to micronutrients stress (deficiency and toxicity) as well as Aluminium stress (toxicity). Proteins were extracted from rice tissue at 4, 6, 8, and 10 day after stress treatment and recovery. The proteins were then acetone precipitated before separation by one dimensional gel electrophoresis. Several protein bands like 257 kDa, 242 kDa, 225 kDa, 210 kDa, 153 kDa, 139kDa, 125 kDa, 101 kDa, 91 kDa, 78 kDa, 64 kDa, and 39 kDa, were found to induced or suppressed by the treatments. The identities of the altered stress proteins can possibly be worked out by obtaining information on the amino acid sequences of the short peptides of these proteins. Proteome analysis can thus distinguish differences in the timing and amount of protein expression altered by micronutrients stress (deficiency and toxicity) as well as Aluminium stress (toxicity).ThesisItem Open Access MEDIA MANIPULATION FOR IN VITRO PLANT REGENERATION IN PEPPERMINT (Mentha piperita L.)(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2007) SINGH, PRAVIN KUMAR; Saxena, AnitaIn Jharkhand, it is grown to some extent in Chotanagpur, Ranchi and Santhal Pargana regions. Its essential oil is called peppermint oil and contains menthol, menthofuron, methone, iso-methone iso-menthol, neomenthol, neo-isomenthol etc. Another important substance is rosmarinic acid. It is an ester of caffeic acid and 3, 4 dihydroxy phenylacetic acid possessing antioxidant and antimicrobial activities. A lot of complex medicines and phyto-preparations are made of mint and menthol. Peppermint oil also has other applications for e.g. as a flavouring agent in confectionary items, alcoholic drinks, dental creams, mouth washes, soaps, chewing gums, candies etc. Its essential oils also have antibacterial and antifungal properties. The present study was undertaken to standardize protocol for in vitro regeneration of this crop. Surface sterilized shoot tips and nodes were inoculated on Murashige and Skoog’s (M.S) medium either supplemented with hormones i.e. 6-Benzylamino purine (B.A.P) and 6, Furfurylamino purine (Kn) (single or incombination) or without hormones. Maximum bud breaking was obtained in B.A.P 1.0 mg/l. In B.A.P 1.0 mg/l and 2.5 mg/l maximum shoot multiplication was observed in case of nodes and shoot tips. Maximum root induction were found in B.A.P 1.0 mg/l. Rooted plants were transferred for hardening. The survival percentage was 90 per cent during hardening.
