Institute of Applied Biosciences, CERTH, Thermi, 570 01, Thessaloniki, Greece.
Perrotis College, American Farm School, 570 01, Thessaloniki, Greece.
Biochem Genet. 2019 Dec;57(6):747-766. doi: 10.1007/s10528-019-09920-6. Epub 2019 Apr 17.
Beans are one of the most important staple crops in the world. Runner bean (Phaseolus coccineus L.) is a small-scale agriculture crop compared to common bean (Phaseolusvulgaris). Beans have been introduced to Europe from the Central America to Europe and since then they have been scattered to different geographical regions. This has resulted in the generation of numerous local cultivars and landraces with distinguished characters and adaptive potential. To identify and characterize the underlying genomic variation of two very closely related runner bean cultivars, we performed RNA-Seq with de novo transcriptome assembly in two landraces of P. coccineus, 'Gigantes' and 'Elephantes' phenotypically distinct, differing in seed size and shape. The cleaned reads generated 37,379 and 37,774 transcripts for 'Gigantes' and 'Elephantes,' respectively. A total of 1896 DEGs were identified between the two cultivars, 1248 upregulated in 'Elephantes' and 648 upregulated in 'Gigantes.' A significant upregulation of defense-related genes was observed in 'Elephantes,' among those, numerous members of the AP2-EREBP, WRKY, NAC, and bHLH transcription factor families. In total, 3956 and 4322 SSRs were identified in 'Gigantes' and 'Elephantes,' respectively. Trinucleotide repeats were the most dominant repeat motif, accounting for 41.9% in 'Gigantes' and 40.1% in 'Elephantes' of the SSRs identified, followed by dinucleotide repeats (29.1% in both cultivars). Additionally, 19,281 putative SNPs were identified, among those 3161 were non-synonymous, thus having potential functional implications. High-confidence non-synonymous SNPs were successfully validated with an HRM assay, which can be directly adopted for P. coccineus molecular breeding. These results significantly expand the number of polymorphic markers within P. coccineus genus, enabling the robust identification of runner bean cultivars, the construction of high-resolution genetic maps, potentiating genome-wide association studies. They finally contribute to the genetic reservoir for the improvement of the closely related and intercrossable Phaseolus vulgaris.
豆类是世界上最重要的主要粮食作物之一。与普通菜豆相比,红花菜豆(Phaseolus coccineus L.)是一种小规模农业作物。红花菜豆从中美洲引入欧洲,此后已散布到不同的地理区域。这导致了许多具有独特特征和适应潜力的本地品种和地方品种的产生。为了鉴定和描述两个非常密切相关的红花菜豆品种的潜在基因组变异,我们对两个红花菜豆地方品种“Gigantes”和“Elephantes”进行了 RNA-Seq 分析,并从头组装转录组。这两个品种表型不同,种子大小和形状不同。对于“Gigantes”和“Elephantes”,分别生成了 37,379 和 37,774 个转录本。在这两个品种之间鉴定了 1896 个差异表达基因(DEGs),其中 1248 个在“Elephantes”中上调,648 个在“Gigantes”中上调。在“Elephantes”中观察到防御相关基因的显著上调,其中包括 AP2-EREBP、WRKY、NAC 和 bHLH 转录因子家族的许多成员。在“Gigantes”和“Elephantes”中分别鉴定了 3956 和 4322 个 SSRs。三核苷酸重复是最主要的重复基序,在“Gigantes”中占 SSRs 的 41.9%,在“Elephantes”中占 40.1%,其次是二核苷酸重复(在两个品种中均占 29.1%)。此外,还鉴定了 19,281 个假定的 SNPs,其中 3161 个是非同义的,因此具有潜在的功能意义。通过 HRM 分析成功验证了高可信度的非同义 SNPs,可直接应用于红花菜豆分子育种。这些结果显著增加了红花菜豆属内的多态性标记数量,使红花菜豆品种的鉴定更加可靠,高分辨率遗传图谱的构建,以及全基因组关联研究的开展成为可能。它们最终为相关且可杂交的普通菜豆的改良提供了遗传资源。