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[转基因番茄植株中花分生组织的异常发育不依赖于编码防御相关病程相关蛋白和抗菌肽的基因的表达]

[Abnormal floral meristem development in transgenic tomato plants do not depend on the expression of genes encoding defense-related PR-proteins and antimicrobial peptides].

作者信息

Khaliluev M R, Chaban I A, Kononenko N V, Baranova E N, Dolgov S V, Kharchenko P N, Poliakov V Iu

出版信息

Ontogenez. 2014 Jan-Feb;45(1):28-41.

Abstract

In this study, the morphological and cytoembryological analyses of the tomato plants transformed with the genes encoding chitin-binding proteins (ac and RS-intron-Shir) from Amaranthus caudatus L. andA. retroflexus L., respectively, as well as the gene amp2 encoding hevein-like antimicrobial peptides from Stellaria media L., have been performed. The transgenic lines were adapted to soil and grown the greenhouse. The analysis of putative transgenic tomato plants revealed several lines that did not differ phenotypically from the wild type plants and three lines with disruption in differentiation of the inflorescence shoot and the flower, as well as the fruit formation (modified plants of each line were transformed with a single gene as noted before). Abnormalities in the development of the generative organs were maintained for at least six vegetative generations. These transgenic plants were shown to be defective in the mail gametophyte formation, fertilization, and, consequently, led to parthenocarpic fruits. The detailed analysis of growing ovules in the abnormal transgenic plants showed that the replacement tissue was formed and proliferated instead of unfertilized embryo sac. The structure of the replacement tissue differed from both embryonic and endosperm tissue of the normal ovule. The formation of the replacement tissue occurred due to continuing proliferation of the endothelial cells that lost their ability for differentiation. The final step in the development of the replacement tissue was its death, which resulted in the cell lysis. The expression of the genes used was confirmed by RT-PCR in all three lines with abnormal phenotype, as well as in several lines that did not phenotypically differ from the untransformed control. This suggests that abnormalities in the organs of the generative sphere in the transgenic plants do not depend on the expression of the foreign genes that were introduced in the tomato genome. Here, we argue that agrobacterial transformation affects, directly or indirectly, expression of genes encoding for transcription factors that can activate a gene cascade responsible for the normal plant development.

摘要

在本研究中,分别对用来自尾穗苋和反枝苋的编码几丁质结合蛋白的基因(ac和RS-intron-Shir)以及来自繁缕的编码类橡胶树抗菌肽的基因amp2转化的番茄植株进行了形态学和细胞胚胎学分析。转基因株系适应土壤并在温室中生长。对推定的转基因番茄植株的分析显示,有几个株系在表型上与野生型植株没有差异,还有三个株系在花序枝和花的分化以及果实形成方面出现异常(如前所述,每个株系的改良植株用单个基因进行转化)。生殖器官发育异常至少持续了六个营养世代。这些转基因植株在雄配子体形成、受精方面存在缺陷,因此导致了单性结实果实。对异常转基因植株中正在发育的胚珠的详细分析表明,未受精的胚囊被替代组织所形成和增殖所取代。替代组织的结构与正常胚珠的胚和胚乳组织均不同。替代组织的形成是由于失去分化能力的内皮层细胞持续增殖所致。替代组织发育的最后一步是其死亡,这导致了细胞裂解。通过RT-PCR在所有三个具有异常表型的株系以及几个在表型上与未转化对照无差异的株系中证实了所用基因的表达。这表明转基因植株生殖领域器官的异常并不取决于导入番茄基因组中的外源基因的表达。在此,我们认为农杆菌转化直接或间接影响编码转录因子的基因的表达,这些转录因子可激活负责正常植物发育的基因级联反应。

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