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Abstract
Plantation white sugar factories in Bangladesh are battling with many problems in recent times. Shortage of quality raw material (i.e. sugarcane) is one of the major problems that paralyse Bangladeshi sugar industry. Investigation is necessary to determine why sugarcane as a cash crop is no longer viable to growers. It is of prime importance to quickly find a solution that satisfies both the growers and mill management. Introduction of a feature rich variety of sugarcane to the growers is a promising solution. Introduction of a sugarcane variety to growers that is disease, draught, bug resistant and obtain high yield/unit area may encourage growers but high sucrose content in sugarcane is the primary concern for mill management. Tissue and Cell culture can play a vital role to cost effectively produce large number of disease free seed of a suitable sugarcane variety. Tissue culture is a modern process and extensive research are been being done on this novel process.
Plant tissue culture may offer certain advantages over traditional methods including: [1] The production of exact copies of plants that produce particularly good flowers, fruits, or have other desirable traits. To quickly produce mature plants. The production of multiples of plants in the absence of seeds or necessary pollinators to produce seeds. The regeneration of whole plants from plant cells that have been genetically modified. The production of plants in sterile containers that allows them to be moved with greatly reduced chances of transmitting diseases, pests, and pathogens. The production of plants from seeds that otherwise have very low chances of germinating and growing, i.e.: orchids and nepenthes. To clean particular plants of viral and other infections and to quickly multiply these plants as 'cleaned stock' for horticulture and agriculture.
The list above mentions certain advantages of plant tissue culture over traditional methods of propagation. So it is obvious that sugarcane growers will prefer tissue cultured seed over traditional one. So multiplication of a sugarcane variety with desirable feature by means of tissue culture may be the first step to address the problem of insufficient supply of quality sugarcane to sugar mills.
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Scope of research
A quick internet search indicated that sugarcane tissue culture research is carried out in different countries including India, Pakistan, The Philippines & Brazil etc. In our sub-continent India & Pakistan have some success story regarding tissue culture of sugarcane. Considering the similarities between Climate Condition of sub-continental countries Indian & Pakistani research were given priority. Internet search also indicated that in Bangladesh very few researches have been carried out on sugarcane tissue culture. [2]
References
[1]http://en.wikipedia.org/wiki/Plant_tissue_culture [2]http://www.baptcb.org/ptc/search.asp?YEAR=sugarcane&B1=Search
Tissue culture
Tissue culture is the cloning of plants from mother plant. Tissues from mother plant are taken out and grown in nutrient media following the protocol. A protocol for a particular variety is a detailed procedure to multiply it. Development of protocol is an iterative and research oriented process. The process involves selection of a mother plant, developing the mother plant into plantlets under controlled conditions, development of roots of these plantlets and finally hardening the plant in the greenhouse.
8. Reduce lodging which will also decrease the infestation of insects and pests. 9. Lower production cost due to high sugar and cane yield per unit area. 10. Vigorous growth and development results greater disease resistance. 11. Availability of high quality vigorous and disease free seed at affordable price round the year. 12 Every bud sprouts at least thrice.
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The lab is promoting commercial cultivation of the following variety of tissue culture sugarcane:
Cultivar C0 86032
Characters It is an improved variety with sucrose content as high as18-21%. It gives about 20-25 tillers/plant. It is a comparatively newer variety developed in 2000 and now gaining popularity due to high yield & sucrose content. It is a comparatively newer variety & now gaining popularity due to high yield & sucrose content.
released by Agriculture
C0 91010 C094012
Variety Name
CP-43-33 CP- 77-400 CP 81-1435 ABT super BF - 162 SPSG - 26 SPF - 234 BL - 4 T - 10
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Recently released varieties of sugarcane by Sugarcane Breeding Institute, Coimbatore, India [7]
Time Period Sub-Tropical Zone 1980s 1990s Co 1148, Co 1158, Co 7717, Co 7314 Co 1148, Co 89003
2000s
Co 740, Co 62175, Co 6304, Co 7219, Co 7704, Co 7527, Co 7508, Co 7504, Co 8011, Co 8014, Co 8021, Co 8208, Co 8362, Co 8371, Co 8338, Co 85004, Co 86032, Co 85019, Co 86249, Co 97009 Co 89003, Co 98014, Co 86032, Co 99004, Co 94012, Co Co 0238, Co 0118 2001-13, Co 2001-15
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lux light intensity. The incubation temperature was 26C 1C with 16 hour light and 8 hour dark period in every 24 hour cycle. First sub-culturing was done after five weeks and rest sub-culturing after two weeks. During each sub-culturing all dead or dis-coloured or vitrified shoots were removed. Hardening was carried out in glass house under natural light conditions.
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Results
Shoot formation from apical meristem: The criterion of good growth for newly formed shoots from apical meristem was based on the production of broad and dark green colored leaves, healthy stems and number of small germinating buds at the base of stem. From Table 1a it is evident that in CP 77400 best results for shoot formation were obtained in AM2 medium (MS medium containing 1.5 mg/l of BAP). In this medium all explants showed shoot proliferation response within 10 days with maximum number of 1.8 shoots per explant (Fig. 2 a). By increasing the concentration of BAP, frequency of shoot proliferation was decreased and time taken for shoot formation was also delayed, In case of BL-4, shoot formation response varied from 9 to 13.5 days. Best response was obtained in AM6 medium (BAP 0.5 mg/l + Kinetin 0.25 mg/l). All explants showed 100% shoot formation within 10 days with 1.7 shoots per explant (Table 1b, Fig. 2b). AM1 and AM8 media also showed hundred percent shoot formation but time taken was more and the number of shoots per explants were less. All other combination did not prove good for shoot formation in BL-4 (Table 1b). In the present investigation shoot apical meristem of different sizes ranging from 0.5-5 mm was used. As shown in Table 2, time for shoot formation was increased by decreasing the size of meristem. Maximum rate of survival was achieved when meristem of 3 mm size was used. This size exhibited 100% survival with 90% regeneration potential within 12 days of inoculation. For shoot formation both solid and liquid media were used. Best results were obtained on media solidified with Phytagel at 3.0 mg/l. Multiple shoot formation: After 45 weeks of shoot growth, actively growing shoots were transferred to fresh medium in jars for further growth and
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proliferation. Both solid and liquid media were tested. Best results for shoot multiplication were obtained in liquid medium. Proliferation of shoot started and during secondary proliferation stage, lateral shoots developed from the base of newly initiated shoot. As a result a dense mass of shoots (25-30) was developed in each culture jar (Fig. 3a and b). After 20 days these bunches were further sub-divided in bunches containing 4-5 shoots and were transferred into fresh medium in jars. In this way shoot multiplication was maintained for several passages by regular transfer to fresh medium. The best shoot multiplication response in CP 77400 was obtained in SM1 medium i.e. MS medium containing 1.0 mg/l BAP (Fig. 4a). In this medium 29 shoots were obtained after four weeks of sub-culturing. Addition of kinetin and GA did not show any support to shoot multiplication in CP 77400. In case of BL-4, best shoot multiplication was achieved in SM5 medium i.e., MS media containing 0.25 mg/l BAP and Kin each (Fig. 4b). From Table 3 it is evident that in both the varieties rate of shoot multiplication increased by decreasing the concentration of BAP. It was also observed that shoot multiplication response was enhanced in liquid medium while solid medium delayed shoot multiplication response.
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References
[1]http://www.vsisugar.com/india/agriculture_divisions/tissue_culture/micropropagationtechnology.htm [2] http://www.vsisugar.com/india/products/tissue_plantlets.htm [3]http://www.pakissan.com/english/newtech/sugarcan.shtml [4]http://www.advanceagriculturalpractice.in/w/index.php/Sugarcane_Cultivation_Package [5]http://www.ablr.co.in/productions2.html [6] http://www.pakissan.com/english/newtech/sugarcan.shtml [7] http://www.sugarcane.res.in/index.php/research/popular-varieties?start=1 [8] http://www.pakbs.org/pjbot/PDFs/40(1)/PJB40(1)139.pdf
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