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葡萄柚种子无菌苗组培快繁体系的建立(PDF)

《广西植物》[ISSN:1000-3142/CN:45-1134/Q]

期数:
2015年06期
页码:
891-898
栏目:
研究论文
出版日期:
2015-11-20

文章信息/Info

Title:
Establishment of rapid propagation for sterile seedlings of grapefruit by tissue culture
文章编号:
1000-3142(2015)06-0891-08
作者:
叶维雁 郭晓月 刘惠民* 王连春 刘 鹏 吴海波
西南林业大学 国家林业局西南地区生物多样性保育重点实验室, 昆明 650224
Author(s):
YE Wei-Yan GUO Xiao-Yue LIU Hui-Min* WANG Lian-Chun LIU Peng WU Hai-Bo
Key laboratory of Biodiversity Conservation in Southwest China, State Forestry Administration, Southwest Forestry University, Kunming 650224, China
关键词:
葡萄柚 种子 植物生长调节剂 初代培养 继代培养
Keywords:
grapefruit seed plant growth regulator primary culture subculture
分类号:
S666.3
DOI:
10.11931/guihaia.gxzw201411013
文献标识码:
A
摘要:
以葡萄柚种子在无菌条件下萌发的幼苗作为外植体供体,子叶切断作为丛生芽初代诱导培养的接种材料,以MT作为基本培养基,通过调节不同植物生长调节剂及其浓度组合、不同质量浓度的蔗糖,选择最佳初代培养基、继代培养基和生根培养基,对葡萄柚组培快繁技术体系进行研究。结果表明:(1)葡萄柚种子经预处理后,先用75%酒精表面灭菌15 s,再用0.1% HgCl2浸泡20 min的消毒效果最好,污染率为18.33%,萌发率为89.91%;(2)初代培养的最适培养基为MT+1.0 mg·L-16-BA(6-苄氨基腺嘌呤)+0.2 mg·L-1IBA(吲哚丁酸)+0.2 mg·L-1GA(赤霉素)+蔗糖40 g·L-1,腋芽诱导率较高,为93.33%;(3)浓度为0.2 mg·L-1的GA能提高初代培养的腋芽诱导率,但还达不到显著水平;(4)继代培养的最适培养基为MT+0.5 mg·L-16-BA+0.2 mg·L-1IBA+0.2 mg·L-1GA+蔗糖40 g·L-1,丛生芽增殖系数达到4.25,芽深绿色,茎粗壮,节间较长,生长旺盛;(5)浓度为0.2 mg·L-1的GA能显著提高继代培养的增殖系数;(6)最适合继代培养的蔗糖浓度为40 g·L-1,丛生芽长势极好;(7)最佳生根培养基为1/2 MT+NAA 0.2 mg/L+蔗糖40 g·L-1+AC 0.1%,生根率达68.89%,根粗壮;(8)生根苗移栽30 d后成活率在75%以上。该研究建立了葡萄柚的组织培养快繁技术体系,为葡萄柚的规模化生产提供了可行的技术依据。
Abstract:
The most appropriate sterilization method for tissue culture of grapefruit’s seeds was selected, the seedlings sprouted from grapefruit’s seeds under sterile conditions acted as explant donors, the cotyledon cuts were used as the inoculation material of the clustered shoots’ primary induction culture, the best primary culture medium, subculture culture medium and rooting medium were selected by adjusting the concentrations of sucrose, combinations of different plant growth regulators and their concentrations in the MT medium, and the tissue culture and rapid propagation technique system of grapefruit were studied. The results were as follows:(1)Rapidly immersed with 75% alcohol for 15 s plus 0.1% HgCl2 for 20 min after the pretreatment was the best disinfection method for grapefruit seeds with the contamination rate of 18.33%, and the germination rate of 89.91%;(2)The best medium for the primary culture was MT+1.0 mg·L-16-BA(6-Benzylaminopurine)+0.2 mg·L-1IBA(Indole-3-Butytric acid)+0.2 mg·L-1GA(Gibberellic acid)+sucrose 40 g·L-1, the axillary shoot induction rate was up to 93.33%;(3)GA the concentration of which was 0.2 mg·L-1 could improve the axillary shoot induction rate of primary culture, but couldn’t reach significant level;(4)MT+0.5 mg·L-16-BA+0.2 mg·L-1IBA+0.2 mg·L-1GA+sucrose 40 g·L-1 was the most suitable for the subculture with the multiplication coefficient of 4.25, the shoots were dark green with vigorous growth, the stems were thick and internodes were relatively long;(5)GA the concentration of which was 0.2 mg·L-1 could significantly improve the multiplication coefficient of subculture;(6)The most suitable concentration of sucrose for the subculture was 40 g·L-1, the clustered shoots’ growth was excellent;(7)The best rooting medium was 1/2MT+NAA 0.2 mg·L-1+sucrose 40 g·L-1+AC 0.1%+sucrose 40 g·L-1 with the rooting rate of 68.89%, the roots were very sturdy;(8)The transplant survival rate of rooting plantlets was up to 75% after being transplanted for 30 d. Through the experiment the tissue culture and rapid propagation technique system of grapefruit was established, which would provided a feasible technical basis for the mass production of grapefruit.

参考文献/References

Chen SC(陈世昌). 2011. Plant Tissue Culture(植物组织培养)[M]. Beijing(北京): Higher Education Press(高等教育出版社): 33 Chen ZX(陈泽雄), Liu YQ(刘奕清), Lou J(娄 娟). 2007. Improvement of shoot-tip micrografting through adventitious bud induction from Citrus stem explants(利用柑橘茎段诱导不定芽改进微芽嫁接技术的研究)[J]. J Southwest Chin Norm Univ:Nat Sci Ed(西南师范大学学报·自然科学版), 32(1): 57-61
Dong GF(董高峰), Huang T(黄涛), Li GG(李耿光), et al. 2001. Studies on the tube-graftion and micro-propagation of mature shatian pomelo(成年沙田柚树试管嫁接及微繁殖的研究)[J]. Ecol Sci(生态科学), 20(3):20-25
Guo LR(郭林榕), Chen WG(陈文光), Xiong YM(熊月明). 2002. Marketing and development prospects of grapefruit(葡萄柚的产销及发展前景)[J]. Fujian Fruits(福建果树), 3: 34-35
He H(贺红), Pan RC(潘瑞炽), He YW(何亚文), et al. 1997. A study on the tissue culture and plant regeneration of Citrus reticulata cv. Tankan(蕉柑组织培养与植株再生的研究)[J]. J S Chin Norm Univ: Nat Sci Ed(华南师范大学学报·自然科学版), 4: 63-66
Huang JQ, Yin LY, Yang XH,et al. 2005. In vitro plant regeneration from the mature tissue of navel orange(Citrus sinensis L. Osbeck)by direct organogenesis[J]. Agric Sci Chin, 4(3): 236-240
Hong L(洪柳), Liu YZ(刘永忠),Deng XX(邓秀新). 2005. Obtaining of ponkan(Citrus reticulate blanco)tetraploid by culturing embryos of mature seeds(椪柑成熟种子胚培养获得四倍体植株)[J]. Acta Hortic Sin(园艺学报), 32(4): 688-690
Jin JT(金建涛), Lai ZX(赖钟雄), Liu SC(刘生财), et al. 2014. Optimization of plantlet regeneration system and in vitro conservation of Fortunella crassifolia cv. Youxijingan(尤溪金柑离体再生体系优化及试管苗保存)[J]. J Fujian Agric For Univ:Nat Sci Ed(福建农林大学学报·自然科学版), 43(3): 256-262
Jiang L(姜玲), Wan SY(万蜀渊), Wang YH(王映红), et al. 1995. Improvement on the shoot-tip grafting method(柑橘茎尖嫁接操作方法的改进及研究)[J]. J Huazhong Agric Univ(华中农业大学学报), 14(4): 381-385
Liang CF(梁称福). 2005. Literature review of research and application on plant tissue culture(植物组织培养研究进展与应用概况)[J]. Nonwd For Res(经济林研究), 23(4): 99-105 Li JM(李浚明), Zhu DY(朱登云). 2005. Plant Tissue Culture Tutorial(植物组织培养教程)][M]. 3rd Ed(第3版). Beijing(北京): China Agricultural University Press(中国农业大学出版社): 20
Li K(李凯), Chen XT(陈雄庭), Hong L(洪磊), et al. 2011. Improving on mature-type Citrus grandis stem culture in vitro(化州橘红成年态茎段离体培养的改良)[J]. J Trop Organ(热带生物学报), 2(1): 42-45
Li L(李丽), Luo JQ(罗君琴), Xu JG(徐建国), et al. 2011. Investigation of techniques of local characteristic citrus varieties shoot-tip micrografting for virus-free seedling(地方特色柑桔品种茎尖微芽嫁接脱毒育苗技术初探)[J]. Xiandai Hortic(现代园艺), 2: 3+46
Li L(李丽), Luo JQ(罗君琴), Xu JG(徐建国), et al. 2011. Preliminary report of grapefruit shoot-tip micrografting for virus-free seedling(葡萄柚茎尖微芽嫁接脱毒成苗技术研究初报)[J]. Zhejiang Citrus (浙江柑橘), 28(3): 12-13
Lu RS(陆荣生), Han ML(韩美丽), Huo XJ(霍秀娟), et al. 2013. Study on regeneration system of adventitious bud from grapefruit(葡萄柚不定芽再生体系建立研究)[J]. Acta Agric Jiangxi(江西农业学报), 25(6): 35-38
Liu WL(刘文龙), Li JJ(李进进), Chen SY(陈升云), et al. 2009. Discussion on orange micro-shoots grafting in tube(柑橘试管内微芽嫁接方法探讨)[J]. J Guangdong Ind Techn Coll(广东轻工职业技术学院学报), 8(3): 14-16+20 Li Y(李悦), Hou BB(侯滨滨), Jing BY(静宝元). 2010. The antibacterial activity study of grapefruit essential oil(葡萄柚精油抑菌活性的研究)[J]. Food Res Devel(食品研究与开发), 31(11): 237-240
Randall PN, Terence JE. 2011. Mixture screening and mixture-amount designs to determine plant growth regulator effects on shoot regeneration from grapefruit(Citrus paradise macf.)epicotyls[J]. In Vitro Cell Dev Biol-Plant, 47(6): 682-694
Rehana S, Alam MS, Islam KS,et al. 2013. Influence of growth regulators on shoot proliferation and plantlet production from shoot tips of banana[J]. Prog Agric, 20(1): 9-16
Wang CX(王彩霞). 2009a. Effect of plant growth regulators on the morphogenesis of three citrus species(植物生长调节剂对3个柑橘品种形态建成的影响)[J]. Chin Agric Sci Bull(中国农学通报), 25(2):16-19
Wang CX(王彩霞). 2009b. Study on in vitro stem culture of adult citrus(柑橘成年态茎段培养技术体系研究)[J]. Guangdong Agric Sci(广东农业科学), 1: 47-49
Wu HB(吴海波), Liu LM(刘惠民). 2005. Progress of grapefruit cultivation and research(葡萄柚的栽培及研究概况)[J]. Nonwd For Res (经济林研究), 23(1): 69-73
Wang RX(王任翔), Gao CW(高成伟), Li JR(李洁荣), et al. 2003. Tissue culture of Citrus cv. Olinda in vitro(欧林达夏橙组织培养研究)[J]. J Guangxi Norm Univ:Nat Sci Ed(广西师范大学学报·自然科学版), 21(2): 71-74
Wang YY(王玉英), Gao X(高新). 2006. Technical Manual of Plant Tissue Culture(植物组织培养技术手册)[M]. Beijing(北京): The JinDun Publishing House(金盾出版社): 43-44
Wang ZC(王子成), Deng XX(邓秀新). 2002. Cryopreservation of Citrus protoplasts(柑橘原生质体的超低温保存)[J]. J Henan Univ:Nat Sci Ed(河南大学学报·自然科学版), 32(3): 38-40
Wang ZC(王子成), Li ZA(李忠爱), Deng XX(邓秀新). 2005. Studies on the methods of mature Citrus stems disinfection(柑橘成年态茎段外植体消毒方法研究)[J]. J Henan Univ:Nat Sci Ed)(河南大学学报·自然科学版), 35(2): 57-60
Yan CJ(颜昌敬). 1990. Plant Tissue Culture Manual(植物组织培养手册)[M]. Shanghai(上海):Shanghai Science and Technology Press(上海科学技术出版社): 56-79
Yang LZ(杨连珍). 1996. Summary of grapefruit(葡萄柚概述)[J]. Chin J Trop Agric(热带作物研究), 25(2): 67-71
Ye YM(叶荫民). 1997. Grapefruit(Citrus paradisi Macf.)and development prospects in China(葡萄柚(Citrus paradisi Macf.)及其在我国的发展前景)[J]. S Chin Fruits(中国南方果树), 26(5): 7-9
Yang ZN, Ingelbrecht IL, Louzada E,et al. 2000. Agrobacterium-mediated transformation of the commercially important grapefruit cultivar Rio Red(Citrus paradise Macf.)[J]. Plant Cell Reports, 19: 1 203-1 211
Zhou HX(周海霞). 2010. Different virus elimination methods in species variation of Citrus siensis Osbeck cv. Hongjiang and the effect of the virus of liberobacter asiaticum(不同脱毒方法对红江橙种性变异及脱黄龙病毒效果的影响)[D]. Zhanjiang(湛江): Guangdong Ocean University(广东海洋大学) Zhang JY(张家银), Guo C(郭琛), Xie YM(谢玉明), et al. 2008. In vitro system for the regeneration of mature intermodal stem segments of ponkan(Citrus reticulate blanco)(椪柑成年态节间茎段再生体系的建立)[J]. J Hunan Agric Univ:Nat Sci Ed(湖南农业大学学报·自然科学版), 34(2): 182-185

备注/Memo

备注/Memo:
收稿日期: 2014-11-10修回日期: 2015-03-12
基金项目: 国家国际科技合作专项项目(2014DFA31060)
作者简介: 叶维雁(1988-),男,广西合浦县人,在读硕士研究生,主要从事果树组织培养研究,(Email)flyingywy@163.com。 *通讯作者: 刘惠民,博士,教授,现在主要从事经济林栽培与利用等研究工作,(Email)hmliu@swfu.edu.cn。
更新日期/Last Update: 2015-11-20