Zhang Yao, Liu Xi, Li Yuying, Liu Xiongfang, Ma Hong, Qu Suping, Li Zhenghong
Institute of Highland Forest Science, Chinese Academy of Forestry, Kunming 650224, China.
College of Forestry, Nanjing Forestry University, Nanjing 210037, China.
Plants (Basel). 2022 Apr 29;11(9):1204. doi: 10.3390/plants11091204.
(Rubiaceae), an evergreen shrub or small tree, is endemic to China and confined to Nujiang Prefecture, Yunnan Province. This plant is of high ornamental value owing to its attractive pink flowers, sweet fragrance, and long flowering period. Due to the influence of climate change and human factors, the distribution range of has exhibited a significant shrinking trend, and it has become a vulnerable species that is in urgent need of conservation and rational utilization research. In this study, the flower transcriptome sequencing of was conducted using an Illumina HiSeq platform. We designed and developed a series of EST-SSR primers based on the flower transcriptome data of . The results showed that 98,389 unigenes were obtained from the flower transcriptome, all of which were aligned with sequences in public databases. Nr, Nt, Pfam, KOG/COG, Swiss-Prot, KEGG, and GO annotated 31,859, 13,853, 22,684, 10,947, 21,416, 9722, and 23,390 unigenes, respectively. The MISA (Microsatellite) tool was used to identify SSR loci from all unigenes, and a total of 15,384 SSRs were identified. Repeat motifs were given priority with mononucleotides, dinucleotides, and trinucleotides. The 81 primer pairs were synthesized randomly, of which 44 pairs showed effective amplification. A total of 17 primers showed stable amplification, and rich polymorphism was observed in 6 populations. We concluded via genetic diversity analysis that the average effective number of alleles (Ne), Shannon's information index (I), and polymorphism information content (PIC) were 1.925, 0.837, and 0.403, respectively. In conclusion, 17 EST-SSR primers can be used for subsequent population genetic diversity analysis and molecular-marker-assisted breeding, which is of great significance for formulating resource conservation and utilization strategies for .
茜草科,一种常绿灌木或小乔木,是中国特有的,仅分布于云南省怒江州。这种植物因其迷人的粉色花朵、甜美的香气和较长的花期而具有很高的观赏价值。由于气候变化和人为因素的影响,其分布范围呈现出显著的缩小趋势,已成为急需进行保护和合理利用研究的易危物种。在本研究中,利用Illumina HiSeq平台对其进行了花转录组测序。我们基于其花转录组数据设计并开发了一系列EST-SSR引物。结果表明,从其花转录组中获得了98389个单基因,所有这些单基因均与公共数据库中的序列进行了比对。Nr、Nt、Pfam、KOG/COG、Swiss-Prot、KEGG和GO分别注释了31859、13853、22684、10947、21416、9722和23390个单基因。使用MISA(微卫星)工具从所有单基因中鉴定SSR位点,共鉴定出15384个SSR。重复基序以单核苷酸、二核苷酸和三核苷酸优先。随机合成了81对引物,其中44对显示出有效扩增。共有17对引物显示出稳定扩增,在6个种群中观察到丰富的多态性。通过遗传多样性分析我们得出结论,平均有效等位基因数(Ne)、香农信息指数(I)和多态性信息含量(PIC)分别为1.925、0.837和0.403。总之,17对EST-SSR引物可用于后续的种群遗传多样性分析和分子标记辅助育种,这对于制定其资源保护和利用策略具有重要意义。