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高密度遗传图谱为菊苣核雄性不育(NMS1)和孢子体自交不亲和(S 座位)相关基因座定位

High-density genetic maps for loci involved in nuclear male sterility (NMS1) and sporophytic self-incompatibility (S-locus) in chicory (Cichorium intybus L., Asteraceae).

机构信息

Université de Lille, UMR INRA-Lille 1 1281, Stress Abiotiques et Différenciation des Végétaux Cultivés, Villeneuve d'Ascq, France.

出版信息

Theor Appl Genet. 2013 Aug;126(8):2103-21. doi: 10.1007/s00122-013-2122-9. Epub 2013 May 21.

Abstract

High-density genetic maps were constructed for loci involved in nuclear male sterility (NMS1-locus) and sporophytic self-incompatibility (S-locus) in chicory (Cichorium intybus L.). The mapping population consisted of 389 F1' individuals derived from a cross between two plants, K28 (male-sterile) and K59 (pollen-fertile), both heterozygous at the S-locus. This F1' mapping population segregated for both male sterility (MS) and strong self-incompatibility (SI) phenotypes. Phenotyping F1' individuals for MS allowed us to map the NMS1-locus to linkage group (LG) 5, while controlled diallel and factorial crosses to identify compatible/incompatible phenotypes mapped the S-locus to LG2. To increase the density of markers around these loci, bulked segregant analysis was used. Bulks and parental plants K28 and K59 were screened using amplified fragment length polymorphism (AFLP) analysis, with a complete set of 256 primer combinations of EcoRI-ANN and MseI-CNN. A total of 31,000 fragments were generated, of which 2,350 showed polymorphism between K59 and K28. Thirteen AFLP markers were identified close to the NMS1-locus and six in the vicinity of the S-locus. From these AFLP markers, eight were transformed into sequence-characterized amplified region (SCAR) markers and of these five showed co-dominant polymorphism. The chromosomal regions containing the NMS1-locus and the S-locus were each confined to a region of 0.8 cM. In addition, we mapped genes encoding proteins similar to S-receptor kinase, the female determinant of sporophytic SI in the Brasicaceae, and also markers in the vicinity of the putative S-locus of sunflower, but none of these genes or markers mapped close to the chicory S-locus.

摘要

高密度遗传图谱被构建用于研究菊苣中的核雄性不育(NMS1 基因座)和孢子体自交不亲和(S 基因座)相关基因。该作图群体由来自于两个植株 K28(雄性不育)和 K59(花粉可育)的杂交后代 389 个 F1'个体组成,这两个植株在 S 基因座均为杂合子。该 F1'作图群体在雄性不育(MS)和强自交不亲和(SI)表型上分离。对 F1'个体进行 MS 表型分析将 NMS1 基因座定位到连锁群(LG)5 上,而通过控制的自交和析因杂交鉴定可亲和/不可亲和表型将 S 基因座定位到 LG2 上。为了增加这些基因座周围标记的密度,采用了混池分离分析法。使用扩增片段长度多态性(AFLP)分析对混池和亲本植株 K28 和 K59 进行筛选,使用了一套完整的 256 对 EcoRI-ANN 和 MseI-CNN 引物组合。共产生了 31000 个片段,其中 2350 个在 K59 和 K28 之间表现出多态性。在 NMS1 基因座附近鉴定出 13 个 AFLP 标记,在 S 基因座附近鉴定出 6 个 AFLP 标记。从这些 AFLP 标记中,有 8 个转化为序列特征扩增区域(SCAR)标记,其中 5 个标记表现出共显性多态性。包含 NMS1 基因座和 S 基因座的染色体区域各自局限在 0.8 cM 的区域内。此外,我们还定位了编码与 Brassicaceae 中的孢子体自交不亲和的 S 受体激酶的雌性决定因子相似的蛋白的基因,以及向日葵中假定 S 基因座附近的标记,但这些基因或标记都没有靠近菊苣的 S 基因座。

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