中国常用橡胶树品种鉴定方法研究

Rapid Identification of Rubber Tree Varieties Commonly used in China

橡胶树(Hevea brasiliensis)产生的胶乳是天然橡胶的最重要来源,不同品种产生胶乳的能力不同,但不同的橡胶树品种形态差异小,难以区分。利用简单的方法区分不同的橡胶树品种具有重要的意义。以天然橡胶生物合成相关的7个基因家族中的目的基因及启动子序列为模板,设计基因特异的PCR引物16对,分别扩增21个橡胶树栽培品种的基因组DNA,扩增条带的多态性可将19个品种区分开(除了云研77-4和南华1号外)。再利用橡胶小粒子编码蛋白基因(SRPP2)特异的引物,扩增云研77-4和南华1号的SRPP2基因,PCR反应产物直接测序,通过单核苷酸水平上的差异可准确的将二者区分开。本研究通过简单的PCR扩增及其扩增产物的直接测序,建立了一种稳定性好、简单快速、经济高效的鉴定橡胶树品种的方法。

The latex of the rubber tree （Hevea brasiliensis） is the most important source of natural rubber. The ability of latex production and adaptability of different rubber tree clones varied greatly, however, it is difficult to distinguish clones based on morphological traits, which is an important task in field work. In this study, sixteen pairs of gene specific primers were designed basing on gene and promoter sequences of genes related to natural rubber biosynthesis, these primer pairs were used respectively to amplify the genomic DNA of 21 rubber tree clones widely grown in China. The results showed that banding profile of these primer pairs could distinguish the 19 rubber tree clones, except Yunyan77-4 and Nanhual. PCR amplification products using gene （SRPP2 encoding small rubber particle protein）specific primers were used for sequencing analysis, these two Clones could be distinguished based on single nucleotide differences. In this study, a simple, stable and cost effective method using PCR band profile and PCR product sequencing to identify rubber tree clones was established.