摘要
试管苗作为马铃薯良种繁育的核心材料,其快繁壮苗是马铃薯种薯生产的基础环节。只有在有限时间内培育出根系发达、茎杆粗壮、有效茎节数多的试管苗,才更有利于新品种优质试管薯的诱导,对促进新品种在生产中尽快推广应用有着极其重要的现实意义。
本试验采用青海省农林科学院作物所马铃薯研究室育成的2个马铃薯品种,高淀粉型品种青薯2号和食品加工型品种青薯6号,研究了植物生长调节剂对马铃薯试管苗生长的影响,建立不同品种试管苗的优化培养体系,为马铃薯快繁提供理论依据,主要试验结论如下:
1.采用五因素(2,4-D、6-BA、NAA、KT、GA)四水平正交试验设计,研究了生长调节剂种类和浓度对两个品种试管苗株高、叶片数的影响。结果表明:不同生长调节剂和浓度对试管苗株高、叶片数有显著影响,说明不同品种的遗传基础不同,因此对于不同品种试管苗的培育应采用不同的培养基。
2.差异显著性检验表明:2,4-D对试管苗株高的作用最显著,5种激素对试管苗株高的影响顺序为:2,4-D>NAA >6-BA> GA> KT,适合株高生长的最佳培养基为MS基本培养基中附加2,4-D0.5 mg/L,6-BA1.0 mg/L,KT0.9 mg/L,GA6.5 mg/L;NAA对试管苗叶片生长的作用最显著,5种激素对试管苗叶片生长的影响顺序为:NAA > KT>6-BA> 2,4-D>GA,适合叶片生长的最佳培养基为MS基本培养基中附加NAA0.1mg/L,6-BA1.0 mg/L,KT0.1 mg/L,GA3.0 mg/L。
3.筛选出了两个品种试管苗培养基的最佳生长调节剂配方:适合青薯2号试管苗生长的最佳培养基为MS基本培养基添加2,4-D0.5 mg/L,6-BA1.0mg/L,KT0.9 mg/L,GA6.5 mg/L;适合青薯6号试管苗生长的最佳培养基为MS基本培养基添加2,4-D2.5mg/L,6-BA2.0mg/L,KT0.1 mg/L,GA10.0 mg/L。
4.运用饱和D最优设计方法,建立了不同植物生长调节剂、不同外植体数对试管苗生长的数学模型。最佳培养方案:试管苗最佳的外植体个数为7—9个,2,4-c最佳浓度为1.32mg/L—2.514mg/L;6-BA最佳浓度1.05mg/L—1.95mg/L;NAA最佳浓度0.165mg/L—0.335mg/L;KT最佳浓度0.35mg/L—0.65 mg/L;GA最佳浓度3.5mg/L—6.5mg/L.
As basic stock of potato breeding and multiplication, to obtain vigorous seedlings in vitro was a crucial step. How to obtain vigorous seedlings with health roots, thick stems and more functional nodes would be beneficial for inducing micro tubers. It was also a practical mean to boost popularization of new varieties.
Two varieties breed by laboratory of potato, institute of crop, agricultural academy of Qinghai province were applied to this test that are Qingshu 2 which was the high starch variety and Qingshu 6 which was the processing food varieties. In order to provide theoretical basis for rapid propagation, the effects of plant growth regulators on growth of in vitro potato platelet was studied. And the optimized culture system of different varieties of in vitro potato platelet was established. The major experimental conclusions were as follows:
1. The effects of kinds and concentration of plant growth regulators on plant height and leaf number of two varieties in vitro potato platelet was studied by orthogonal test design L16(45). The five-factor were 2,4-D,6-BA,NAA,KT and GA. The result showed that significant effect of different kinds and concentration of plant growth regulators on plant height and leaf number, which meant genetic background among varieties tested were different, and separate medium should be adopted for various varieties.
2. The test of the significance of difference was showed that 2,4-D had the most prominent effect on plant height followed by NAA, 6-BA, GA and KT. The best medium for lengthwise growth was MS with 2,4-D0.5 mg/L, 6-BA1.0 mg/L, KT0.9 mg/L and GA6.5 mg/L; NAA had the most prominent effect on leaf numbers followed by KT, 6-BA, 2,4-D and GA. The best medium for leaf growth was MS with NAA0.1mg/L, 6-BA1.0 mg/L, KT0.1 mg/L and GA3.0 mg/L.
3. It was screened out the optimum formula of plant growth regulators for culture medium of two varieties in vitro potato platelet. The best medium for growth of qingshu 2 was MS with 2,4-D0.5 mg/L, 6-BA1.0mg/L,KT0.9 mg/L and GA6.5 mg/L the growth; The best medium for growth of qingshu 6 was MS with 2,4-D2.5mg/L,6-BA2.0mg/L,KT0.1 mg/L and GA10.0 mg/L.
4. The mathematical model of different plant growth regulators and different explants on growth of in vitro potato platelet were established by D-saturating optimum design. Those were the optimum formula: the best explants number was 7—9; the best concentration of 2,4-D was 1.32mg/L—2.514mg/L; the best concentration of 6-BA was 1.05mg/L—1.95mg/L;the best concentration of NAA was 0.165mg/L—0.335mg/L; the best concentration of KT was 0.35mg/L—0.65mg/L,and the best concentration of GA was 3.5mg/L—6.5mg/L.
引文
1. Ancora G , A Sonnino. In vitro induction of mutation in potato[M]. In Bajaj YPS (ed) Biotechnology in Agriculture and Forestry 3: Potato, Springer Verlag Berlin Heidelberg, Germany. IV 12,1987.
2. Hawkes J G. The potato evolution, biodiversity and genetic resources [M]. Belhaven press(a division of Pinter Publishers), London,1990.
3. 陈萌山.中国马铃薯产业发展现状及展望[M].第五届世界马铃薯大会文集,中国昆明,2004,81-82.
4. 张树丛,张东霖.马铃薯退化及解决途径[J].宁夏科技,1996(3):30-33.
5. 苏琴英,刘万福.马铃薯退化原因及防止退化措施[J].甘肃农村科技,2003,(4):30-34.
6. Luis F Salazar.马铃薯病毒及其防治[M].阎文昭,张勇飞等译.中国农业科技出版社,2001,183-184.
7. 王小菁,李玲主编.植物生长调节剂在植物组织培养中的应用[M].化学工业出版社,2003.
8. 邵莉楣,孟小雄.植物生长调节剂应用手册[M].金盾出版社,2000.
9. 韩德元.植物生长调节剂原理与应用[M].北京科学技术出版社,1997.
10. Weaver,R.J.. Plant Growth Substances in Agriculture[M].Freeman & Co.,Sanfrancisco Calif,1972.
11. Luckwill L.C.. Growth Regulators in Crop Production[M].Edward Arnojd, London,1981.
12. Robetts,J.A., Hooley R. Plant Growth Regulators[M].Blackie Glasgow ,London,1988.
13. Miller P R. The use of plant growth regulation in micro propagation of slow growing potato cultivars [J]. Potato Research,1985,28(2):479-486.
14. 贾长盛等.植物生长调节剂对下寨 65 试管苗生长的影响[M].中国马铃薯学术研讨文集,黑龙江科学技术出版社,1996,144-148.
15. 韩善华,郑国昌.马铃薯幼茎(芽)愈伤组织的诱导和植株再生[J].实验生物学报,1982,15(4):10-15.
16. 唐洪明,王林萍等.内蒙古西部区马铃薯脱毒种薯快速繁育的研究[J].中国马铃薯,1997,2:2-9.
17. 崔凯荣,邢更生等.植物激素对体细胞胚胎发生的诱导与调节[J].遗传,2002,22(5):349-354.
18. Acram T Prakash K, Prakash L. Plant growth regulators. In vitro plant breeding [J]. Food products press,2001,129-131.
19. 姜应德.马铃薯试管苗壮苗培育初探[M].植物组织培养与脱毒快繁技术,中国科学技术出版社,2001,270-272.
20. Tovar R,Estrada et al. Induction of in vitro potato tubers [M]. CIP Circularl3 (4) 1-4. Internal potatoCenter, Lima, peru,1985.
21. 张维本,王兴杰等.马铃薯脱毒试管苗简易培养基研究初报[J].中国马铃薯,1997,2:29-30.
22. 马春红,崔四平等.不同激素浓度及抗生素对马铃薯脱毒试管苗的影响[J].河北农业科学 ,1999,3(2): 8-10.
23. 祝红艺,柴岩等.KT 与 NAA 对马铃薯组培苗生长的影响[J].河北农业科学,2000,4(2):7-9.
24. 刘小凤,吴云锋等.不同浓度 NAA 和 KT 对马铃薯组培苗的影响及方程模型[J].西北农业学报,2005,14(6):106-108.
25. 李凤云,韩丽颖.外源激素对马铃薯脱毒试管苗微繁的影响[J].中国马铃薯,2002,16(4): 214-216.
26. 李文刚.马铃薯脱毒种薯微型化繁育推广体系研究进展[M].中国马铃薯研究与产业开发,哈尔滨工程大学出版社,2003,108-112.
27. 仲乃琴,王一航等.碳源浓度对不同光源培养的马铃薯脱毒试管苗生长的影响[J].中国马铃薯,2000,14(2):73-75.
28. 辛国斌,陈远达.马铃薯脱毒试管苗液体静止培养过程中玻璃化的预防及壮苗措施[J].中国马铃薯,2003,17(2):95-96.
29. 徐传朝,张夏菊等.活性炭对马铃薯试管苗生长和微型薯诱导影响研究[J].中国农业科技导报,2003,5(5):107-108.
30. 方志明.加入抗生素培养基对马铃薯试管苗的影响[J].中国马铃薯,2001,15(2):92.
31. 王春.医用抗生素在马铃薯组织培养中的抑菌效应研究[J].甘肃农业科技,2004,10:15-16.
32. 祁彦丰,王萃莲等.用三种不同水质配制培养基对马铃薯脱毒试管苗的影响[J].中国马铃薯,2000, 14(3):173.
33. 齐恩芳,仲乃琴等.不同培养方式和成分对马铃薯脱毒试管苗生长的影响[J].中国马铃薯,2000,14(1):18-19.
34. 郝文胜,赵永秀等.我国马铃薯茎尖培养脱毒和脱毒试管苗微繁研究进展[J].内蒙古农业科技,(内蒙古农业职业教育专集),2001,27-33.
35. 颜谦,黄萍等.马铃薯脱毒试管苗高效生产技术[J].种子杂志,2004,23(9):82-83.
36. 吴 凌娟 , 张 雅 奎. 马 铃 薯 茎尖 脱 毒 技 术 在 大 兴 安 岭 种 薯 生 产 上 的 应 用 [J]. 中国 马 铃薯,1998,3:35-37.
37. 滕 伟 丽 , 杨 琦 . 倍 力 凝 在 马 铃 薯 脱 毒 试 管 苗 生 产 中 的 应 用 研 究 [J]. 马 铃 薯 杂志,1999,13(4):202-204.
38. 郝云凤,李可伟等.脱毒马铃薯快繁培养基的不同支持物[J].马铃薯杂志,2005,19(1):42-43.
39. 谢庆华,昊毅效等.固定物对马铃薯脱毒试管苗生长的影响[J].中国马铃薯,2001,15(1):20-21.
40. 罗智敏,王炳君等.卡拉胶和琼脂为固定物对马铃薯脱毒组培苗生长的影响[J].马铃薯杂志,1997, 11(2):73-74.
41. 马淑珍.MS 全脱水培养基培养马铃薯脱毒试管苗效果初报[J].马铃薯杂志,1999,13(4):219-220.
42. 林丛发,魏泽平等.马铃薯脱毒试管苗繁育及脱毒种薯生产技术[J].中国马铃薯,2000,4:225-226.
43. 连勇.马铃薯脱毒快繁及工厂化生产技术[M].植物组织培养与脱毒快繁技术,中国科学技术出版社, 2001:264-269.
44. 李文刚,梁东超等.马铃薯脱毒微型薯工厂化繁育技术研究[J].中国马铃薯,2001,15 (5):278-280.
45. 孔繁春,李文刚等.不同培养方式和光照对马铃薯脱毒试管苗微繁的影响[J].马铃薯杂志,1999,13(1):20-22.
46. 高慧君,陈涛.马铃薯脱毒试管苗自然光照培养研究[J].马铃薯杂志,1996,10(1):29-31.
47. 陈凯,刘颖等.培养基与光照强度对马铃薯脱毒试管苗组培快繁的影响[J].安徽农业科学, 2005,33(9):16-28.
48. Moe R, Field T et al. Stem elongation and Keeping quality in poinsettia as affected by temperature and supplementary lighting [J]. Sei Hortic,1992,50:127-136.
49. Ahn YulKyun, Na Seung Yong et al. The effects of photoperiods on in vitro microtuberization in potato (Solarium tuberosum L.) [J]. Journal of the Korean Society for Horticultural Science,2002,43(3):301-305.
50. 牛爱国,侯丽娟等.马铃薯组培苗液体静置培养微繁技术研究[J].马铃薯杂志,1999,13(2):35-38.
51. 白艳菊,李学湛等.优质、低成本工厂化生产马铃薯脱毒试管苗[J].中国农学通报,2001,17 (2):82-83.
52. 叶彦,缪树华.长期继代培养马铃薯愈伤的植株再生[J].应用与环境生物学报,1995,1(1):26-33.
53. 增田芳雄,胜见允行等.植物激素[M].科学出版社,1976.
54. Nora A,Choveaux. The Effect of 1-NapHthaleneacetic Acid on the Endogenous Cytokinin Content of Aseptically Cultured Bark Segments of Salix babylonica [J]. Plant Cell PHysiol,1981,22(7):1207-1214.
55. Abou Mandoura A, Hartung W J. The Effect of Abscisic Acid and Increased Osmotic Potential of theMedia on Growth and Root Regeneration of Zea mays Callus [J].Plant PHysiol,1986,22:139-145.
56. Hirishik Amada, Hiroshi Harada. Changes in the Endogenous Leves and Effects of Abscisic Acid during Somatic Embryogenesis of Daucus carota L [J]. Plant Cell PHysoil,1981,22(8):1423-1429.
57. Luckwill L C .Growth Regulators in Crop Production[M]. Edward Arnojd,London,1981.
58. 张永成.饱和-D 最优设计方法在农业试验中的应用[J].中国马铃薯,1997,03:117-176.
59. 徐福祥.马铃薯脱毒种薯繁育体系的建立及其技术措施[J].农业科技通讯,1992,1:10-12.
60. 张维本,王兴杰等.马铃薯脱毒试管苗简易培养基研究初报[J].中国马铃薯,1997,2:29-30.
61. 王巧玲,李淑芳等.马铃薯脱毒试管苗壮苗培育初探[J].中国马铃薯,1999,3:27-28.
62. 金顺福,姜成模.培育健壮马铃薯试管苗试验[J].马铃薯杂志,1995,9(3):139-143.
63. 马伟青,王培伦等.培养基中氮磷钾对马铃薯试管苗生长的影响[J].山东农业科学,1999,4: 36-37.
64. 王付欣,铁双贵等.马铃薯试管苗快繁技术研究[J].河南农业科学,2003,(7):44-47.