[1]曲玲,焦恩宁,李彦龙,等. 枸杞抗炭疽病菌毒素愈伤组织变异体的离体筛选及其防御酶活性研究[J].西北林学院学报,2015,30(03):81-88.[doi:doi:10.3969/j.issn.1001-7461.2015.03.15]
　QU Ling,JIAO En-ning,LI Yan-long,et al. Selection of Resistant Calli of Lycium barbarum to Colletotrichum gloeosporioides in vitro and Its Changes in Activity of Defensive Enzymes[J].JOURNAL OF NORTHWEST FORESTRY UNIVERSITY,2015,30(03):81-88.[doi:doi:10.3969/j.issn.1001-7461.2015.03.15]
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 枸杞抗炭疽病菌毒素愈伤组织变异体的离体筛选及其防御酶活性研究()
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《西北林学院学报》[ISSN:1001-7461/CN:61-1202/S]

卷:
第30卷
期数:
2015年03期
页码:
81-88
栏目:
出版日期:
2015-05-31

文章信息/Info

Title:
 Selection of Resistant Calli of Lycium barbarum to Colletotrichum gloeosporioides in vitro and Its Changes in Activity of Defensive Enzymes
文章编号:
1001-7461(2015)03-0081-08
作者:
 曲玲; 焦恩宁; 李彦龙; 戴国礼; 石志刚; 安巍; 刘兰英; 曹有龙*
 （宁夏农林科学院 国家枸杞工程技术研究中心，宁夏 银川 750002）
Author(s):
 QU Ling;  JIAO En-ning;  LI Yan-long;  DAI Guo-li;  SHI Zhi-gang;  AN Wei;  LIU Lan-ying;  CAO You-long*
 (National Engineering Research Center of Wolfberry, Ningxia Academy of Agricultural and Forestry Sciences, Yinchuan, Ningxia 750002, China)
关键词:
枸杞; 炭疽病; 粗毒素; 离体筛选; 愈伤组织变异体; 诱变; 防御酶活性
Keywords:
Lycium barbarum;  Colletotrichum gloeosporioides;  crude toxin;  in vitro selection;  resistant callus variant;  inductive variation;  activity of defensive enzyme
分类号:
S722.35
DOI:
doi:10.3969/j.issn.1001-7461.2015.03.15
文献标志码:
A
摘要:
 以枸杞炭疽病菌粗毒素液作为选择剂，将体细胞无性系变异与辐射诱变相结合，对宁夏枸杞主栽品种“宁杞1号”的叶片胚性愈伤组织进行辐射诱变和筛选，以期获得稳定的抗炭疽病细胞变异系。结果表明，以半致死剂量的60Co γ对枸杞叶片诱导出的胚型愈伤组织进行辐照处理，将其恢复增殖后，采用多步正选择法结合加压与去压交替的筛选方法，筛选到了抗性愈伤组织变异体，其在不同粗毒素浓度胁迫下，其鲜重增长均高于对照组，而且在离开选择因子继代培养4代后，仍保持获得的抗性。分析抗性系与抗病相关的防御酶的活性表明，无论有无粗毒素胁迫，抗性愈伤组织变异体的PAL、POD和PPO酶活性均高于对照愈伤组织；在粗毒素胁迫下，随着粗毒素浓度的增大，3种酶活性基本呈先升高后降低的趋势，抗性变异系3种酶活性的上升幅度较对照组要大，而PAL和POD活性的下降速度较对照亲本系小，说明抗性变异系通过增强与抗病性相关酶的活性，来加强同植物防卫有关的物质代谢。
Abstract:
 For obtaining stable cell mutant of Lycium barbarum. resistant to Colletotrichum gloeosporioides, the disease resistant mutant selection system was established by the integral application of theγ-ray irradiation and somaclonal variation techniques. The results showed some stable cell mutants resistant to the toxin of C. gloeosporioides were obtained after the calli induced with the leaves of Ningqi No.1 (L.barbarum cv. Ningqi 1) were treated by the dose value of LD50 of 60Co γ-ray irradiation and then selected by using multi-step screening system and interval selected method. The fresh weight increment of the resistant lines was higher than that of controls at different concentrations of crude toxin, and they could remain resistance to toxin after they were subcultured four times successively apart from selective factor. Furthermore, changes of activity of defensive enzymes (PAL, POD and PPO) in mutant and original callus were investigated after treated by different concentrations of crude toxin. The results demonstrated that the activity of three disease resistance-related enzymes in resistant calli was higher than that of parent calli whether or not treated with crude toxin. As the concentration of crude toxin increased, the activity of three defensive enzymes both increased then decreased, the range of the activity variation of three enzymes in variant cell lines was lower than those of the parent.

参考文献/References:

 [1]王荔，罗文富，杨艳琼，等. 运用致病毒素筛选抗烟草黑胫病细胞突变体I抗毒素愈伤组织的有效筛选[J].云南农业大学学报, 1999, 14(1):16-21.
WANG L, LUO W F, YANG Y Q, et al. Using phytotoxin to select mutants resistant to tobacco Phytophthora parasitia var. nicotianae Ⅰ. selection of callus resistant to phytotoxin[J]. Journal of Yunnan Agricultural University, 1999, 14(1):16-21. (in Chinese )
[2]王荔, 杨艳琼, 杨德, 等. 运用致病毒素筛选抗烟草黑胫病细胞突变体II抗毒素愈伤组织的再生植株及其后代的抗病性鉴定[J]. 云南农业大学学报, 1999, 14(3):273-278. 
WANG L, YANG Y Q, YANG D, et al. Selection of mutants resistance to tobacco Phytophthora parasitica var. nicotianae using phytotoxin II, resistance to black shank fungus inoculation of regeneration plants and their progenies of toxin-resistant callus[J]. Journal of Yunnan Agricultural University, 1999, 14(3):273-278. (in Chinese )
[3]郭丽娟，胡启德，康绍兰，等. 诱发玉米抗小斑病突变体的研IV. 从玉米单倍体胚性细胞无性系筛选抗玉米小斑病突变体[J]. 遗传学报, 1987, 14(5):355-362.
GUO L J, HU Q D, KANG S L, et al. Studies on induction of Helminthosporium maydis resistance mutation in Zea mays IV. the screening of disease-resistant mutation from haploid embryonic cell clone in Zea mays[J]. Acta Genetic Sinica, 1987, 14(5):355-362. (in Chinese)
[4]曾列先，徐羡民，伍尚忠，等. 以水稻白叶枯病原菌培养过滤液离体筛选再生植株后代的抗病性研究[J]. 植物保护学报，1992，19(3)：231-235.
ZENG L X, XU X M, WU S Z, et al. Studies on disease resistance for the progeny of rice plants regeneration with tissue culture treated by the culture filtrates of Xanthomomas campestris pv. oryzea[J]. Acta Phytophylactca Sinica, 1992, 19(3):231-235. (in Chinese )
[5]曲玲，石志刚，焦恩宁，等．枸杞炭疽病菌滤液对枸杞愈伤组织生长及分化的影响[J]. 北方园艺, 2012(23):112-115.
[6]KOUKOL J, CONN E E. The metabolism of aromatic and properties of the phenylalanine deaminase of Hordeum vulgare[J]. Journal of Biological Chemistry, 1961, 236(10):2692-2698.
[7]陈建勋. 植物生理学实验指导[M]. 广州：华南理工大学出版社，2002：120-121.
[8]李靖，利容千，袁文静. 黄瓜感染霜霉病菌叶片中一些酶活性的变化[J]．植物病理学报，1991, 21(4)：277-282.
LI J, LI R Q, YUAN W J. On the change of enzyme activities of cucumber leaf infected by Pseudoperonospora cubensis(Berk.et Curt .) Rostov[J]. Acta Phytopathologica Sinica, 1991, 21(4):277-282. (in Chinese)
[9]朱至清. 体细胞无性系变异与植物改良[C]// 魏建昆.农业高新技术论. 北京：科学技术文献出版社，1993:129-135.

[10]蔡其华，高明慰，梁竹青．离体诱变改良杂交水稻的研究[J]. 核农学报, 1990,4(3):169-174. 
CAI Q H, GAO M W, LIANG Z Q. In vitro culture mutagenesis for hybrid rice improvement[J]. Journal of Nuclear Agricultural Sciences, 1990, 4(3):169-174. (in Chinese)
[11]高明尉，梁竹青，成雄鹰．诱变处理对小麦体细胞组织离体培养的效应[J]．中国农业科学，1987,20(1):20-3l .
[12]郑企成，朱耀兰，陈文华，等．小麦体细胞无性系变异中γ射线辐照效果[J]．植物学通报，1991，8(Supp.1)：78-82. 
[13]王敬驹. 细胞突变体筛选[C]// 孙敬三.植物细胞工程实验技术. 北京：科学出版社，1995:127.
[14]李社荣，李安生，张敬，等．小麦抗赤霉病突变体的离体筛选及其生化变化的研究[J]．核农学报，1995，16(3)：158-161.
[15]彭明生，郭兆年．苹果抗斑点落叶病突变体的细胞筛选[J]. 果树科学，1992，9(4)：213-217.
[16]刘海瑞，许文耀，林成辉. 利用枯萎病菌粗毒素筛选香蕉抗性突变体[J]．亚热带农业研究，2007，3(3)：211-214. 
[17]林丛发，钟爱清，翁琳琳，等. 利用叶斑病菌粗毒素离体筛选太子参抗叶斑病突变体[J]. 中国农学通报，2010，26(23)：42-47. 
[18]HARTMAN C L, MCCOY T J, KNOUS T R, et al. Selection of alfalfa (Medicago sativa) cell lines and regeneration of plants resistant to the toxin(s) produced by Fusarium oxysporum f. sp. medicaginis[J]. Plant Science Letter, 1984, 84(1):183-194.
[19]李社荣，张敬，李安生，等．小麦抗赤霉菌毒素细胞变异体的筛选及再生植株生化特性分析[J]．农业生物技术学报，1997，5(3)：216-219.
LI S R, ZHANG J, LI A S, et al. Screening of cell lines resistant to Fusarium graminearum and characterization of their regenerated plantlets[J]. Joural of Agricultural Biotechnology, 1997, 5(3):216-219. (in Chinese)
[20]孙祖光．小麦抗根腐病突变体的离体诱变及其抗病机理的研究[M]．北京：原子能出版社，1992.
[21]于翠梅，张月杰，曹萍，等．水稻抗稻瘟病突变体筛选初报[J]．沈阳农业大学学报，2000，31(2)：153-157.
YU C M, ZHANG Y J, CHAO P, et al. Preliminary study on the screening of rice blast-resistant mutant using phytotoxin[J]. Journal of Shenyang Agricultural University, 2000，31(2)：153-157. (in Chinese)
[22]薛应龙，欧阳光察，等．植物抗病的物质代谢基础[C]// 余叔文.植物生理与分子生物学.北京：科学出版社，1998:775-776.
[23]屈雁朋，房玉林，刘延琳，等. 镉胁迫下接种AM真菌对葡萄次生代谢酶活性的影响[J]. 西北林学院学报，2009，24(5)：101-105.
QU Y P, FANG Y L, LIU Y L, et al. Effects of AM fungal on the secondary metabolites of grape under cadmium stress[J]. Journal of Northwest Forestry University, 2009, 24(5):101-105. (in Chinese)
[24]SANCHO M A, FORCHETTI S M, PLIEGO F, et al. Peroxidase activity and isoenzymes in the culture medium of NaCl adapted tomato to suspension cell[J]. Plant Cell, Tissue and Organ Culture, 1996, 44（2）:161-167.
[25]王颖，胡景江，朱玮，等. 杨树溃疡病寄主诱导抗病性的研究[J]. 西北林学院学报，1996，11(1)：1-4. 
WANG Y, HU J J, ZHU W, et al. Induced resistance of poplar to canker[J]. Journal of Northwest Forestry University, 1996, 11(1):1-4. (in Chinese)
[26]王颖，景耀. 植物诱导抗病机理的研究进展[J]. 西北林学院学报，1997，12(3)：52-57. 
WANG Y, JING Y. Advance in the study of the mechanism of plant induced disease resistance[J]. Journal of Northwest Forestry University, 1997, 12(3):52-57. (in Chinese)
[27]辛福梅，张存旭，王红．栎皮栎胚性和非胚性愈伤组织抗氧化物酶的研究[J]．西北林学院学报，2007，22(3)：67-70.
XIN F M, ZHANG C X, WANG H. Antioxides characteristics of embryogenic callus and non-embryogenic callus in Quercus variabilis[J]. Journal of Northwest Forestry University, 2007, 22(3):67-70. (in Chinese)
[28]周自云，胡景江，王俊明．几种激发子对杨树愈伤组织保护酶活性的诱导作用[J]．西北林学院学报，2006，21(1)：43-46.
ZHOU Z Y, HU J J, WANG J M. Reduction by elicitors on four enzymes in poplar callus[J]. Journal of Northwest Forestry University, 2006, 21(1):43-46. (in Chinese)
[29]宋凤鸣，郑重，葛秀春．活性氧及膜脂过氧化在植物-病原菌互作中的作用[J]．植物生理学通迅，1996，32(5)：377-385. 
SONG F M, ZHENG Z, GE X C. Role of active oxygen and membrane lipid peroxidation in plant-pathogen interactions[J]. Plant Physiology Communication, 1996，32(5)：377-385. (in Chinese)
[30]潘瑞炽．植物生理学[M]．北京：人民教育出版社，1979.
[31]杜蕙，王生容．辣椒不同抗病品种感染辣椒疫霉病菌后几种酶活性的变化[J]．农业生物灾害预防与控制研究，1984（3）：278-281. 
[32]杨家书，李舜芳， 吴畏，等．小麦品种对白粉病抗病性与过氧化物酶的关系[J]．植物病理学报，1984，14(4)：235-240.
YANG J S, LI S F, WU W. Varietal resistance of wheat to powdery mildew and its relation to peroxidases[J]. Acta Phytopathologica Sinca, 1984, 14(4):235-240. (in Chinese) 
[33]徐建华，利容千，王建波．黄瓜不同抗病品种感染镰刀菌枯萎病菌后几种酶活性的变化[J]．植物病理学报，1995，25(3)：239-242.
XU J H, LI R Q, WANG L B. Some changes of enzyme activities from susceptible and resistant cucumber cultivars after inoculation with cucumber wilt Fusarium[J]. Acta Phytopathologica Sinica, 1995, 25(3):239-242. (in Chinese)

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备注/Memo

备注/Memo:
 收稿日期：2014-07-07 修回日期：2014-10-16
基金项目：国家自然科学基金(31360167)；宁夏回族自治区自然科学基金(NZ12251)。
作者简介：曲玲，女，硕士，副研究员，研究方向：枸杞抗病性和枸杞生物技术抗病育种。E-mail:cutepearl-1230@163.com
*通信作者：曹有龙，男，博士，研究员，研究方向：枸杞生物技术育种。E-mail:youlongc@hotmail.com
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