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在成熟抑制(rin)番茄成熟突变体中积累模式改变的新型成熟相关mRNA的cDNA克隆与表征

cDNA cloning and characterisation of novel ripening-related mRNAs with altered patterns of accumulation in the ripening inhibitor (rin) tomato ripening mutant.

作者信息

Picton S, Gray J, Barton S, AbuBakar U, Lowe A, Grierson D

机构信息

University of Nottingham, UK.

出版信息

Plant Mol Biol. 1993 Oct;23(1):193-207. doi: 10.1007/BF00021431.

Abstract

A cDNA library produced from mRNA isolated from the pericarp of wild-type tomato fruit (Lycopersicon esculentum Mill. cv Ailsa Craig) at the first visible sign of fruit ripening was differentially screened to identify clones whose homologous mRNAs were present at reduced levels in fruit of the tomato ripening mutant, ripening inhibitor,rin. Five clones were isolated (pERT 1, 10, 13, 14, 15). Accumulation of mRNA homologous to each of these clones increased during the ripening of wild-type fruit and showed reduced accumulation in ripening rin fruit. The levels of three of them (homologous to ERT 1, 13 and 14) were increased by ethylene treatment of the mutant fruit. A further clone, ERT 16 was identified for a mRNA present at a high level in both normal and mutant fruit at early stages of ripening. Database searches revealed no significant homology to the DNA sequence of ERT 14 and 15; however, DNA and derived amino acid sequence of ERT 1 both contain regions of homology with several reported UDP-glucosyl and glucuronosyl transferases (UDPGT) and with a conserved UDPGT motif. A derived amino acid sequence from the ERT 10 cDNA contains a perfect match to a consensus sequence present in a number of dehydrogenases. The ERT 13 DNA sequence has homology with an mRNA present during potato tuberisation. The presence of these mRNAs in tomato fruit is unreported and their role in ripening is unknown. The ERT 16 DNA sequence has homology with a ripening/stress-related cDNA isolated from tomato fruit pericarp.

摘要

从野生型番茄果实(Lycopersicon esculentum Mill. cv Ailsa Craig)果皮中分离出的mRNA构建的cDNA文库,在果实成熟的首个可见迹象时进行差异筛选,以鉴定其同源mRNA在番茄成熟突变体“成熟抑制子”(rin)果实中表达水平降低的克隆。分离出了5个克隆(pERT 1、10、13、14、15)。与这些克隆中每一个同源的mRNA在野生型果实成熟过程中积累增加,而在成熟的rin果实中积累减少。其中3个克隆(与ERT 1、13和14同源)的水平通过对突变体果实进行乙烯处理而升高。还鉴定出了另一个克隆ERT 16,其对应的mRNA在成熟早期的正常果实和突变体果实中均高水平存在。数据库搜索显示ERT 14和15的DNA序列与其他序列无显著同源性;然而,ERT 1的DNA和推导的氨基酸序列均包含与一些已报道的UDP - 葡萄糖基转移酶和葡萄糖醛酸基转移酶(UDPGT)以及一个保守的UDPGT基序的同源区域。ERT 10 cDNA推导的氨基酸序列与多种脱氢酶中存在的共有序列完全匹配。ERT 13的DNA序列与马铃薯块茎形成过程中存在的一种mRNA具有同源性。这些mRNA在番茄果实中的存在尚未见报道,它们在成熟过程中的作用也未知。ERT 16的DNA序列与从番茄果实果皮中分离出的一个与成熟/胁迫相关的cDNA具有同源性。

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