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利用抑制性消减杂交和 cDNA 微阵列技术研究无子温州蜜柑与其有籽野生型在花器官发育过程中的基因表达差异比较转录组分析。

Comparative transcript profiling of gene expression between seedless Ponkan mandarin and its seedy wild type during floral organ development by suppression subtractive hybridization and cDNA microarray.

机构信息

Key Laboratory of Horticultural Plant Biology (Ministry of Education); National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, 430070, China.

出版信息

BMC Genomics. 2012 Aug 16;13:397. doi: 10.1186/1471-2164-13-397.

Abstract

BACKGROUND

Seedlessness is an important agronomic trait for citrus, and male sterility (MS) is one main cause of seedless citrus fruit. However, the molecular mechanism of citrus seedlessness remained not well explored.

RESULTS

An integrative strategy combining suppression subtractive hybridization (SSH) library with cDNA microarray was employed to study the underlying mechanism of seedlessness of a Ponkan mandarin seedless mutant (Citrus reticulata Blanco). Screening with custom microarray, a total of 279 differentially expressed clones were identified, and 133 unigenes (43 contigs and 90 singletons) were obtained after sequencing. Gene Ontology (GO) distribution based on biological process suggested that the majority of differential genes are involved in metabolic process and respond to stimulus and regulation of biology process; based on molecular function they function as DNA/RNA binding or have catalytic activity and oxidoreductase activity. A gene encoding male sterility-like protein was highly up-regulated in the seedless mutant compared with the wild type, while several transcription factors (TFs) such as AP2/EREBP, MYB, WRKY, NAC and C2C2-GATA zinc-finger domain TFs were down-regulated.

CONCLUSION

Our research highlighted some candidate pathways that participated in the citrus male gametophyte development and could be beneficial for seedless citrus breeding in the future.

摘要

背景

无籽性是柑橘的一个重要农艺性状,而雄性不育(MS)是柑橘无核果实的主要原因之一。然而,柑橘无核的分子机制仍未得到很好的探索。

结果

本研究采用抑制性消减杂交(SSH)文库与 cDNA 微阵列相结合的综合策略,研究了温州蜜柑无核突变体(Citrus reticulata Blanco)无核的潜在机制。利用定制的微阵列进行筛选,共鉴定出 279 个差异表达克隆,测序后获得了 133 个基因(43 个串联和 90 个单核苷酸)。基于生物过程的基因本体(GO)分布表明,大多数差异基因参与代谢过程,并对刺激和生物过程的调节做出响应;基于分子功能,它们具有 DNA/RNA 结合或催化活性和氧化还原酶活性。与野生型相比,无核突变体中一个编码雄性不育样蛋白的基因高度上调,而一些转录因子(TFs)如 AP2/EREBP、MYB、WRKY、NAC 和 C2C2-GATA 锌指结构域 TFs 下调。

结论

本研究强调了一些参与柑橘雄配子体发育的候选途径,这对未来的柑橘无核育种可能是有益的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7417/3495689/e8ac9725f4d5/1471-2164-13-397-1.jpg

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