目的 建立不同生態(tài)型天麻Gastrodia elata 葉綠體SSR（chloroplast SSR，cpSSR）分子標(biāo)記鑒定方法，為天麻分子標(biāo)記輔助育種提供理論依據(jù)。方法 利用天麻葉綠體基因組序列獲取cpSSR位點(diǎn)，分析比較位點(diǎn)特征并篩選多態(tài)性cpSSR，通過聚類分析、主成分分析和遺傳結(jié)構(gòu)分析對(duì)不同生態(tài)型天麻遺傳多樣性進(jìn)行分析與鑒定。結(jié)果 天麻葉綠體基因組中篩選到166個(gè)cpSSR位點(diǎn)，其中單核苷酸為主要類型，A/T重復(fù)基序占比最高。篩選得到6對(duì)cpSSR，共擴(kuò)增出18個(gè)條帶，等位基因平均數(shù)（number of allele，N_a）、有效等位基因數(shù)目（effective number of alleles，N_e）、Shannon多樣性指數(shù)（Shannon index，I）、多態(tài)性信息含量指數(shù)（polymorphism information content，PIC）分別為3、2.084 5、0.511 0、0.511 0，表明cpSSR多態(tài)性較高。通過聚類分析、主成分分析和遺傳結(jié)構(gòu)分析均可將不同生態(tài)型天麻分為2個(gè)類群，其中烏天麻、綠天麻和血紅天麻為一類，紅天麻和黃天麻為一類。進(jìn)一步根據(jù)CH15和CH20 2對(duì)cpSSR的電泳特異性條帶，可以將血紅天麻、烏天麻、綠天麻、紅天麻和黃天麻進(jìn)行準(zhǔn)確的鑒定。結(jié)論 利用CH15和CH20兩對(duì)cpSSR鑒定不同生態(tài)型天麻具有效率高、適應(yīng)性廣的優(yōu)點(diǎn)，對(duì)天麻種質(zhì)鑒定具有重要意義。

Objective To establish a chloroplast SSR (cpSSR) molecular marker method for identifying different ecotypes of Gastrodia elata, so as to provide a theoretical foundation for molecular marker-assisted breeding of G. elata.Methods The cpSSR loci were identified from the chloroplast genome sequence of G. elata, followed by the analysis and screening polymorphic cpSSRs. Cluster analysis, principal component, and genetic structure were used to compare and identify the genetic diversity of different ecotypes of G. elata. Results The chloroplast genome of G. elata contained 166 cpSSR loci, with mononucleotide being the most common and A/T repeats motifs being the most frequent. Six pairs cpSSR were selected, which were amplified 18 bands in total. The average number of allele (N_a), effective number of allele (N_e), Shannon index (I), and polymorphism information content (PIC) were 3, 2.084 5, 0.511 0, and 0.511 0, respectively, that indicating high polymorphism in the selected cpSSRs. Genetic diversity analyses, including clustering, principal component analysis, and genetic structure analysis, successfully grouped the G. elata ecotypes into two clusters. One group included the forms of G. elata Bl. f. glauca S. Chow, G. elata Bl. f. viridis Makino, and dark red G. elata Bl., the other one group included G. elata Bl. f. elata and G. elata Bl. f. flavida S. Chow. Using the cpSSR markers CH15 and CH20, these five G. elata forms could be distinguished by specific bands. Conlusion The cpSSR method utilizing CH15 and CH20 for G. elata ecotype identification is efficient and adaptable, which provides a useful tool for germplasm identification of G. elata.

R286.12

國家重點(diǎn)研發(fā)計(jì)劃（2023YFC3503803）;中央本級(jí)重大增減支項(xiàng)目（2060302）;貴科合學(xué)術(shù)新苗[2023]-45