SSAP analysis reveals candidate genes associated with deastringency in persimmon (Diospyros kaki Thunb.) treated with 40 °C water
SSAP analysis reveals candidate genes associated with deastringency in persimmon (Diospyros kaki Thunb.) treated with 40 °C water
复制标题
DOI：
10.1007/s11295-015-0841-6
复制
发表时间：
2015-02
影响因子：
2.4
通讯作者：
C. Guan;L. Chen;Wenxing Chen;Rongli Mo;Qing-lin Zhang;Xiaoyun Du;Jihong Liu;Z. Luo
中科院分区：
文献类型：
--
作者：
C. Guan;L. Chen;Wenxing Chen;Rongli Mo;Qing-lin Zhang;Xiaoyun Du;Jihong Liu;Z. Luo
温水处理是一种简单有效的柿子脱涩方法。在中国授粉恒定和非涩（C-PCNA）品种'鄂柿1号'中的涩可以在40 °C水中快速去除，并且与反转录转座子DkRE 1的激活相结合。本研究采用序列特异性扩增多态性（SSAP）技术，对40 °C水处理后'鄂石1号'柿果实转录水平的变化进行了研究，以探讨与脱涩相关的基因。利用互补DNA（cDNA）-SSAP技术分析了283个可能与‘鄂柿1号’甜柿脱涩有关的差异表达转录衍生片段（TDFs）。总共有116个差异表达的TDFs被注释为具有已知功能的基因，包括4个醛代谢基因和2个果胶相关基因。结果表明，40 °C水处理柿果后，单宁-果胶复合物的形成和果胶酶的促凝作用是导致柿果脱涩的主要原因。值得注意的是，鉴定了三个基因片段，磷酸丙糖异构酶（TDF 175-3）、丙酮酸激酶（TDF 284-2）和新薄荷糖脱氢酶（TDF 238-2），其潜在地参与40 °C水凝固效应。据我们所知，本报告是第一个确定这些基因片段在柿子。将SSAP方法创新性地应用于基于高密度多态性的基因发现。表达分析和转录组数据证实了cDNA-SSAP方法发现新候选基因的有效性和准确性。这些成功分离的基因有助于理解柿子脱涩的机理。
Warm water treatment is a simple and effective deastringency method for the persimmon fruit. The astringency in the ‘Eshi 1’ persimmon, a Chinese pollination-constant and non-astringent (C-PCNA) cultivar, can be quickly removed in 40 °C water coupled with the activation of the retrotransposonDkRE1. In this study, transcriptional level changes in ‘Eshi 1’ persimmon treated with 40 °C water were studied with a sequence-specific amplification polymorphism (SSAP) approach to investigate the genes associated with deastringency. The complementary DNA (cDNA)-SSAP analysis revealed that 283 differentially expressed transcript-derived fragments (TDFs) potentially involved in ‘Eshi 1’ persimmon deastringency were successfully cloned and sequenced. In total, 116 differentially expressed TDFs were annotated as genes with known function, including four aldehyde metabolism genes and two pectin-related genes. Our data suggested that the tannin-pectin complex formation and the aldehyde-mediated coagulation effect contributed to the deastringency of the persimmon fruit treated with 40 °C water. Notably, three gene fragments, triosephosphate isomerase (TDF 175-3), pyruvate kinase (TDF 284-2), and neomenthol dehydrogenase (TDF 238-2), which are potentially involved in the 40 °C water coagulation effect were identified. To our knowledge, this report is the first to identify these gene fragments in the persimmon. The SSAP method was innovatively applied to gene discovery based on high-density polymorphisms. Expression analysis and transcriptome data confirmed the validity and accuracy of the cDNA-SSAP method for discovering new candidate genes. These successfully isolated genes could facilitate understanding of astringency removal in persimmon.