Production and RAPD verification of T0 salt-tolerant alfalfa introducing the total DNA from Rhizophora apiculata by the pollen-tube pathway

Abstract

Abstract: The exogenous total DNA of Rhizophora apiculata was introduced into alfalfa (Medicago sativa) by the pollen-tube pathway. In this study, 1 391 flowers were treated and 894 transformed T₀ seeds were harvested. Twelve T₀ seedlings with improved salt-tolerance were obtained when the T₀ seeds were planted in MS medium containing 225 mmol/L NaCl. RAPD analysis of T₀ seedlings, receptor and donor were carried out. Eight primers which could amplify steady and clear bands were chosen from 55 arbitrary primers tested, and increased band diversity with new bands, specific bands in donor and deletion in receptor. It showed that the exogenous DNA was conformed for integration into the receptors genome, and the improved salt-resistance of T₀ seedlings may be related to the introduction of exogenous genes.

CLC Number:

ZHANG Li-quan, NIU Yi-ding, HAO Jin-feng, HASI Agula. Production and RAPD verification of T₀ salt-tolerant alfalfa introducing the total DNA from Rhizophora apiculata by the pollen-tube pathway[J]. Acta Prataculturae Sinica, 2011, 20(3): 292-297.

References

[1] 赵可夫, 李法曾. 中国盐生植物[M]. 北京: 科学出版社, 1999.
[2] 崔润丽, 刁现民. 植物耐盐相关基因克隆与转化研究进展[J]. 中国生物工程杂志, 2005, 25(8): 25-29.
[3] Mccuc K F, Hanson A D. Drought and salt tolerance: Towards understanding and application[J].Trends in Biotechnology, 2000, 8: 358-362.
[4] Zhou G Y, Weng J, Zeng Y, et al. Introduction of exogenous DNA into cotton embryos[J]. Methods in Enzymology, 1983, 101: 433-481.
[5] 林栖凤, 方清, 邓用川, 等. 耐盐番茄分子育种研究[J]. 生命的化学, 2004, 24: 35-37.
[6] 林栖凤, 邓用川, 李冠一, 等. 外源DNA导入豇豆提高耐盐能力的研究[J]. 生物化学杂志, 1997, 10: 454-455.
[7] 林栖凤, 邓用川, 李冠一, 等. 红树DNA导入茄子获得耐盐性后代的研究[J]. 生物工程进展, 2001, 21(5): 40-44.
[8] 林栖凤, 邓用川, 吴多桂, 等. 耐盐辣椒分子育种[J]. 生物工程进展, 1999, 19(5): 19-24.
[9] 肖军, 张立军, 任志莹, 等. 通过花粉管通道导入外源DNA 获得抗盐番茄新品系[J]. 沈阳农业大学学报, 2008, 39(3): 362-364.
[10] 耿华珠, 吴永敷, 曹致中, 等. 中国苜蓿(第一版)[M]. 北京: 中国农业出版社, 1995.
[11] 马春晖, 夏艳军, 韩军, 等. 不同青贮添加剂对紫花苜蓿青贮品质的影响[J]. 草业学报, 2010, 19(1): 128-133.
[12] 刘青芳, 步怀宇, 赵宇玮, 等. 紫花苜蓿离体培养植株再生及其RAPD 分析[J]. 草业学报, 2006, 15(5): 115-121.
[13] 张改娜, 贾敬芬. 豌豆清蛋白1(AP1)基因的克隆及对苜蓿的转化[J]. 草业学报, 2009, 18(3): 117-125.
[14] Winicov I, Bastola D R. Transgenic overexpression of the transcription factor Alfin1 enhances expression of the endogenous MsPRP2 gene in Alfalfa and improves salinity tolerance of the plants[J]. Plant Physiology, 1999, 120: 473-480.
[15] 燕丽萍, 夏阳, 梁慧敏, 等. 转BADH基因苜蓿T₁代遗传稳定性和抗盐性研究[J]. 草业学报, 2009, 18(6): 65-71.
[16] 吴多桂, 林栖凤, 李冠一. 红树DNA 的CTAB 法提取及其RAPD反应[J]. 中国生物化学及分子生物学报, 1999, 15(1): 67-70.
[17] 王关林, 方宏筠. 植物基因工程(第二版)[M]. 北京: 科学出版社, 2002.
[18] Atienzar F A, Jha A N. The random amplified polymorphic DNA (RAPD) assay and related techniques applied to genotoxicity and carcinogenesis studies: A critical review[J]. Mutation Research, 2006, 613: 76-102.
[19] 王松文, 赵炳然, 邢全华, 等. 野生稻 DNA 片段存在于高世代水稻变异系进一步的分子验证[J]. 中国农业科学, 2006, 39(11): 2170-2177.
[20] 李希臣, 雷勃钧, 卢翠华, 等.早熟大豆外源DNA导入的RAPD分子验证[J]. 大豆科学, 1994, 13(2): 152-156.

Production and RAPD verification of T₀ salt-tolerant alfalfa introducing the total DNA from Rhizophora apiculata by the pollen-tube pathway

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

[1]	XIE Kai-yun,LI Xiang-lin,HE Feng,WAN Li-qiang,WANG Dong,QIN Yan,YU Qun. Response of alfalfa and smooth brome to nitrogen fertilizer in monoculture and mixed grasslands [J]. Acta Prataculturae Sinica, 2014, 23(6): 148-156.
[2]	TIAN Chun-li, JIE Xiao-lei, LIU Yan, LIU Fang, GUO Xiao, HU Hua-feng, LIU Shi-liang. Effects of Se-Zn and fulvic acid combined application on nutrient component and amino acids formation of alfalfa [J]. Acta Prataculturae Sinica, 2014, 23(2): 66-75.
[3]	PENG Lan-qing, LI Xin-yong, QI Xiao, YUE Yan-hong, FAN Shu-gao, LI Shu-cheng, WANG Yan-rong. The relationship of root traits with persistence and biomass in 10 alfalfa varieties [J]. Acta Prataculturae Sinica, 2014, 23(2): 147-153.
[4]	XIA Zeng-run, DU Feng-feng, LI Si, ZHANG Ji-yu, LIU Yong, HUO Ya-xin, KONG Lin-fang. Construction of an EMS induced mutant library and identification of morphological characteristics in Medicago sativa [J]. Acta Prataculturae Sinica, 2014, 23(2): 215-222.
[5]	WU Jing, CAI Hua, BAI Xi, JI Wei, WEI Zheng-wei, TANG Li-li, ZHAO Yang, ZHU Yan-ming. An analysis of salt tolerance of transgenic alfalfa with the GsGST13/SCMRP gene [J]. Acta Prataculturae Sinica, 2014, 23(1): 257-265.
[6]	HAN Hua-wen, SUN Li-na, YAO Tuo, RONG Liang-yan, LIU Qing-hai, LU Hu, MA Hui-ling. Effects of bio-fertilizers with different PGPR strain combinations on yield and quality of alfalfa [J]. Acta Prataculturae Sinica, 2013, 22(5): 104-112.
[7]	XU Yan, WEI Xiao-hong, LI Bing-bing, CAO Li, TANG Zhi-ming. Effects of exogenous nitric oxide on seed germination and seedling oxidative damage in Medicago sativa under NaCl stress [J]. Acta Prataculturae Sinica, 2013, 22(5): 145-154.
[8]	WANG Wei-dong, YANG Pei-zhi, ZHANG Pan, HAN Bo, ZHANG Zhi-qiang, CAO Yu-man, HU Tian-ming. The effect of symbiotic rhizobium on the antioxidative and osmoregulatory capability in alfalfa under salt stress [J]. Acta Prataculturae Sinica, 2013, 22(5): 120-127.
[9]	HUO Ping-hui, LI Jian-feng, SHI Shang-li, ZHANG Shu-qing. Effects of seed ultra-drying storage on growth and resistance of Medicago sativa seedlings affected by neutral salt stress [J]. Acta Prataculturae Sinica, 2013, 22(5): 175-182.
[10]	LIU Jing, CAI Hua, LIU Ying, ZHU Yan-ming, JI Wei, BAI Xi. A study on physiological characteristics and comparison of salt tolerance of two Medicago sativa at the seedling stage [J]. Acta Prataculturae Sinica, 2013, 22(2): 250-256.
[11]	MA Xia, WANG Li-li, LI Wei-jun, SONG Jiang-ping, HE Yuan, LUO Ming. Effects of different nitrogen levels on nitrogen fixation and seed production of alfalfa inoculated with rhizobia [J]. Acta Prataculturae Sinica, 2013, 22(1): 95-102.
[12]	WU Xu-dong, ZHANG Xiao-juan, XIE Ying-zhong, XU Kun, YANG Jing. Vertical distribution characters of soil organic carbon and soil enzyme activity in alfalfa field with different growing years [J]. Acta Prataculturae Sinica, 2013, 22(1): 245-251.
[13]	LIU Yi, ZHANG Chun-mei, XIE Xiao-rong,YAN Fang, HU Dan. Effect of drought stress on polyamine metabolism in the leaves and roots of alfalfa [J]. Acta Prataculturae Sinica, 2012, 21(6): 102-107.
[14]	WANG Xiao-feng, LUO Zhen, LIU Xiao-yan, ZHU Dan, XIA Zhi-qiang, ZHOU Jian-hua, ZHANG Lei. Reparations of Ca²⁺ and P on N-fixation of alfalfa-rhizobia after acid aluminum stress [J]. Acta Prataculturae Sinica, 2012, 21(6): 108-116.
[15]	LI Yan, SUN Yan, YANG Qing-chuan, KANG Jun-mei, ZHANG Tie-jun, FANG Feng. Construction and transformation of an over-expression plasmid of the MsZIP gene from Medicago sativa [J]. Acta Prataculturae Sinica, 2012, 21(6): 182-189.