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本文引用的文献

1
Agrobacterium-mediated transformation of citrange: factors affecting transformation and regeneration.农杆菌介导的枳橙转化:影响转化和再生的因素
Plant Cell Rep. 1998 Dec;18(3-4):271-278. doi: 10.1007/s002990050570.
2
Agrobacterium-mediated in planta genetic transformation of sugarcane setts.农杆菌介导的甘蔗茎段体内遗传转化。
Plant Cell Rep. 2015 Oct;34(10):1835-48. doi: 10.1007/s00299-015-1831-8. Epub 2015 Jul 8.
3
An efficient Agrobacterium-mediated transformation of strawberry cv. Camarosa by a dual plasmid system.通过双质粒系统利用农杆菌介导的高效草莓品种卡玛罗莎转化
Molecules. 2015 Feb 23;20(3):3647-66. doi: 10.3390/molecules20033647.
4
Progress in genetic engineering of peanut (Arachis hypogaea L.)--a review.花生(Arachis hypogaea L.)遗传工程的进展——综述。
Plant Biotechnol J. 2015 Feb;13(2):147-62. doi: 10.1111/pbi.12339.
5
Application of sonication in combination with vacuum infiltration enhances the Agrobacterium-mediated genetic transformation in Indian soybean cultivars.超声处理与真空渗透相结合的应用提高了农杆菌介导的印度大豆品种的遗传转化效率。
Appl Biochem Biotechnol. 2015 Feb;175(4):2266-87. doi: 10.1007/s12010-014-1360-x. Epub 2014 Dec 6.
6
Genetic transformation of green bean callus via Agrobacterium mediated DNA transfer.通过农杆菌介导的 DNA 转移实现绿豆愈伤组织的遗传转化。
Plant Cell Rep. 1993 Jan;12(2):74-9. doi: 10.1007/BF00241938.
7
An efficient in planta transformation of Jatropha curcas (L.) and multiplication of transformed plants through in vivo grafting.麻疯树的高效体内转化及通过体内嫁接繁殖转化植株
Protoplasma. 2014 May;251(3):591-601. doi: 10.1007/s00709-013-0558-z. Epub 2013 Oct 23.
8
Assessment of factors influencing the Agrobacterium-mediated in planta seed transformation of brinjal (Solanum melongena L.).评估影响农杆菌介导的茄子(Solanum melongena L.)体内种子转化的因素。
Appl Biochem Biotechnol. 2013 Sep;171(2):450-68. doi: 10.1007/s12010-013-0359-z. Epub 2013 Jul 14.
9
Agrobacterium tumefaciens-mediated in planta seed transformation strategy in sugarcane.农杆菌介导的甘蔗体内种子转化策略。
Plant Cell Rep. 2013 Oct;32(10):1557-74. doi: 10.1007/s00299-013-1467-5. Epub 2013 Jun 8.
10
Groundnut improvement: use of genetic and genomic tools.花生改良:遗传和基因组工具的利用。
Front Plant Sci. 2013 Feb 25;4:23. doi: 10.3389/fpls.2013.00023. eCollection 2013.

花生[(L.)]的不依赖基因型且增强的植物介导遗传转化

Genotype-independent and enhanced in planta mediated genetic transformation of peanut [ (L.)].

作者信息

Karthik Sivabalan, Pavan Gadamchetty, Sathish Selvam, Siva Ramamoorthy, Kumar Periyasamy Suresh, Manickavasagam Markandan

机构信息

1Department of Biotechnology, Bharathidasan University, Tiruchirappalli, 620024 Tamil Nadu India.

2School of Bio Sciences and Technology, VIT, Vellore, 632014 Tamil Nadu India.

出版信息

3 Biotech. 2018 Apr;8(4):202. doi: 10.1007/s13205-018-1231-1. Epub 2018 Mar 29.

DOI:10.1007/s13205-018-1231-1
PMID:29607283
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5874222/
Abstract

infection and regeneration of the putatively transformed plant from the explant remains arduous for some crop species like peanut. Henceforth, a competent and reproducible in planta genetic transformation protocol is established for peanut cv. CO7 by standardizing various factors such as pre-culture duration, acetosyringone concentration, duration of co-cultivation, sonication and vacuum infiltration. In the present investigation, strain EHA105 harboring the binary vector pCAMBIA1301- was used for transformation. The two-stage selection was carried out using 4 and 250 mg l BASTA to completely eliminate the chimeric and non-transformed plants. The transgene integration into plant genome was evaluated by GUS histochemical assay, polymerase chain reaction (PCR), and Southern blot hybridization. Among the various combinations and concentrations analyzed, highest transformation efficiency was obtained when the 2-day pre-cultured explants were subjected to sonication for 6 min and vacuum infiltrated for 3 min in suspension, and co-cultivated on MS medium supplemented with 150 µM acetosyringone for 3 days. The fidelity of the standardized in planta transformation method was assessed in five peanut cultivars and all the cultivars responded positively with a transformation efficiency ranging from minimum 31.3% (with cv. CO6) to maximum 38.6% (with cv. TMV7). The in planta transformation method optimized in this study could be beneficial to develop superior peanut cultivars with desirable genetic traits.

摘要

对于花生等一些作物品种而言,从外植体诱导出假定转化植株并使其感染和再生仍然很困难。因此,通过对预培养时间、乙酰丁香酮浓度、共培养时间、超声处理和真空渗透等各种因素进行标准化,为花生品种CO7建立了一种高效且可重复的体内遗传转化方案。在本研究中,使用携带双元载体pCAMBIA1301-的EHA105菌株进行转化。采用4和250 mg l的BASTA进行两阶段筛选,以完全消除嵌合体和未转化的植株。通过GUS组织化学分析、聚合酶链反应(PCR)和Southern杂交来评估转基因整合到植物基因组中的情况。在分析的各种组合和浓度中,当将预培养2天的外植体在悬浮液中进行6分钟的超声处理和3分钟的真空渗透,并在添加150 µM乙酰丁香酮的MS培养基上共培养3天时,获得了最高的转化效率。在五个花生品种中评估了标准化体内转化方法的准确性,所有品种均有积极响应,转化效率范围从最低的31.3%(CO6品种)到最高的38.6%(TMV7品种)。本研究中优化的体内转化方法可能有助于培育具有理想遗传性状的优良花生品种。