Development of a protocol for genetic transformation of Malus spp

IF 2.1 4区生物学 Q2 Agricultural and Biological Sciences

Caryologia Pub Date : 2021-12-21 DOI:10.36253/caryologia-1248

F. Martinelli, A. Perrone, A. Dandekar

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引用次数: 0

Abstract

A protocol to produce transgenic shoots of Malus X domestica cv Greensleaves was optimized using two gene constructs previously used to create parthenocarpic tomato, Ino-IaaM and DefH9-IaaM. The aim was to obtain sufficient nº of transgenic shoots for in vitro multiplication, transfer to soil, grafting and testing for parthenocarpy in the next years. We investigated the effects of two modifications of a previous published protocol: 1) co-transformation with an Agrobacterium containing “VIP” genes in the gene construct and 2) two different hormones or hormone combinations. More shoot regeneration was obtained with a combination of three hormones (BA:NAA:TDZ) during co-cultivation instead of IBA and no co-transformation was performed using the VIP gene. For the DefH9-IaaM transgene, 21.04% regeneration was achieved for this treatment instead of 8.95% achieved with “IBA treatment” and 4.42% with the Agrobacterium co-transformation treatment. More shoot regeneration occurred with the combination of three hormones (BA:NAA:TDZ) instead of with only IBA and no co-transformation was performed using VIP gene. Experiments using Ino-IaaM confirmed the results shown for the DefH9-IaaM transgene. The regenerated shoots were multiplied in selective media containing kanamycin and roots were obtained.

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苹果属植物遗传转化方案的建立

利用先前用于单性繁殖番茄的两个基因构建体Ino-IaaM和DefH9-IaaM，优化了国产苹果(Malus X domestica cv Greensleaves)转基因苗的培育方案。目的是获得足够的转基因芽的nº，用于体外繁殖，转移到土壤中，嫁接和未来几年的孤雌性试验。我们研究了对先前发表的方案进行两项修改的效果:1)在基因构建中与含有“VIP”基因的农杆菌共转化;2)两种不同的激素或激素组合。在共培养过程中，3种激素(BA:NAA:TDZ)的组合比IBA的组合能获得更多的芽再生，而VIP基因未进行共转化。对于DefH9-IaaM转基因，该处理的再生率为21.04%，而“IBA处理”的再生率为8.95%，农杆菌共转化处理的再生率为4.42%。3种激素(BA:NAA:TDZ)联合使用比单独使用IBA的再生效果更好，VIP基因未发生共转化。使用Ino-IaaM进行的实验证实了DefH9-IaaM转基因的结果。再生芽在含卡那霉素的选择性培养基中繁殖，获得根。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Caryologia 生物-遗传学

CiteScore

1.60

自引率

23.80%

发文量

审稿时长

12 months

期刊介绍： Caryologia is devoted to the publication of original papers, and occasionally of reviews, about plant, animal and human karyological, cytological, cytogenetic, embryological and ultrastructural studies. Articles about the structure, the organization and the biological events relating to DNA and chromatin organization in eukaryotic cells are considered. Caryologia has a strong tradition in plant and animal cytosystematics and in cytotoxicology. Bioinformatics articles may be considered, but only if they have an emphasis on the relationship between the nucleus and cytoplasm and/or the structural organization of the eukaryotic cell.