The ubiquitin extension protein S27a is differentially expressed in developing flower organs of Thompson seedless versus Thompson seeded grape isogenic clones
The ubiquitin extension protein S27a is differentially expressed in developing flower organs of Thompson seedless versus Thompson seeded grape isogenic clones
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DOI：
10.1007/s00299-009-0715-1
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发表时间：
2009-07-01
影响因子：
6.2
通讯作者：
Perl, Avihai
中科院分区：
文献类型：
--
作者：
Hanania, Uri;Velcheva, Margarita;Perl, Avihai
在葡萄属葡萄中。CV.无籽，受精发生但种子败育，属于种子结实类型。为了克隆在花发育过程中差异表达的转录本，利用两个同基因克隆'Thompson seedless'和'Thompson seeded'，在花序发育的三个阶段（从萌芽到开花前20天）进行抑制性消减杂交。正向和反向消减cDNA文库的差异筛选和测序产生了几个单一的EST。一个差异表达的克隆'汤普森'种子与无籽同基因克隆是泛素延伸蛋白S27 a。原位杂交结果表明，在花发育过程中，该基因在‘Thompson’无核无性系的心皮和子房中的表达显著高于有核无性系。与对照相比，该基因的过表达导致植株再生异常并抑制芽的发育;其在胚性愈伤组织中的沉默诱导细胞坏死和愈伤组织死亡，证明该基因在葡萄发育器官中的紧密调控。S27 a在无核花的心皮和珠被中的过表达可能会干扰这些器官的正常发育，导致胚败育和无核。
In Vitis vinifera L. cv. Thompson Seedless, fertilization occurs but seeds abort, a type of stenospermocarpy. To clone transcripts with differential expression during flower development, suppressive subtractive hybridization was carried out using two isogenic clones 'Thompson seedless' and 'Thompson seeded', at three stages of inflorescence development (from bud break to similar to 20 days prior to anthesis). Differential screening and sequencing of a forward and reverse subtractive cDNA library yielded several singleton ESTs. One differentially expressed clone in 'Thompson' seeded versus seedless isogenic clones was the ubiquitin extension protein S27a. In situ hybridization demonstrated its significantly higher expression in the carpel and ovaries of 'Thompson' seedless versus seeded isogenic clones during flower development. Overexpression of this gene resulted in abnormal plant regeneration and inhibited shoot development compared to controls; its silencing in embryogenic callus induced cell necrosis and callus death, evidencing tight regulation of this gene in developing organs of grape. S27a overexpression in carpels and integuments of the seedless flower may interfere with normal development of these organs, leading to embryo abortion and seedlessness.