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Identifying Differentially Expressed Genes in Pollen from Self-Incompatible Wuzishatangju and Self-Compatible Shatangju Mandarins

机译:从自相矛盾的乌兹沙塘桔和自相适应的沙塘桔普通话中鉴定花粉中差异表达的基因

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摘要

Self-incompatibility (SI) is one of the important factors that can result in seedless fruit in Citrus. However, the molecular mechanism of SI in Citrus is not yet clear. In this study, two suppression subtractive hybridization (SSH) libraries (forward, F and reverse, R) were constructed to isolate differentially expressed genes in pollen from “Wuzishatangju” (SI) and “Shatangju” (self-compatibility, SC) mandarins. Four hundred and sixty-eight differentially expressed cDNA clones from 2077 positive clones were sequenced and identified. Differentially expressed ESTs are possibly involved in the SI reaction of “Wuzishatangju” by regulating pollen development, kinase activity, ubiquitin pathway, pollen-pistil interaction, and calcium ion binding. Twenty five SI candidate genes were obtained, six of which displayed specific expression patterns in various organs and stages after self- and cross-pollination. The expression level of the F-box gene (H304) and S1 (F78) in the pollen of “Wuzishatangju” was 5-fold higher than that in “Shatangju” pollen. The F-box gene, S1, UBE2, UBE3, RNaseHII, and PCP were obviously up-regulated in pistils at 3 d after self-pollination of “Wuzishatangju”, approximately 3-, 2-, 10-, 5-, 5-, and 2-fold higher, respectively than that at the same stage after cross-pollination of “Wuzishatangju” × “Shatangju” pistils. The potential involvement of these genes in the pollen SI reaction of “Wuzishatangju” is discussed.
机译:自交不亲和性(SI)是导致柑橘无核果实的重要因素之一。但是,尚不清楚柑橘中SI的分子机制。在这项研究中,构建了两个抑制消减杂交(SSH)库(正向F和反向R),以从“ Wuzishatangju”(SI)和“ Shatangju”(自相容性,SC)普通话中分离花粉中差异表达的基因。对来自2077个阳性克隆的468个差异表达的cDNA克隆进行了测序和鉴定。差异表达的EST可能通过调节花粉发育,激酶活性,泛素途径,花粉雌蕊相互作用和钙离子结合来参与“乌兹沙汤菊”的SI反应。获得了二十五个SI候选基因,其中六个在自花授粉和交叉授粉后在各个器官和各个阶段显示出特定的表达模式。 F-box基因(H304)和S1(F78)在“五子莎汤菊”花粉中的表达水平是“沙糖菊”花粉中的5倍。 “武则沙棠菊”自花授粉后3 d,雌蕊的F-box基因S1,UBE2,UBE3,RNaseHII和PCP明显上调,大约3-,2-,10-,5-,5-和“武则沙汤菊”ד沙汤菊”雌蕊异花授粉后分别比同期高2倍。讨论了这些基因在“乌兹沙汤菊”花粉SI反应中的潜在参与。

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