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用18对EST-SSR引物对核桃属(Juglans L.)中的6个黑核桃组(Sect.Rhysocayon)样品、14个核桃楸组(Sect.Cardiocaryon)样品、28个核桃组(Sect.Juglans)样品和山核桃属(Carya Nutt)中的4个长山核桃(C.illinoinensis Koch.)样品进行了PCR扩增。在18个EST-SSR位点上共获得110个等位基因,每个位点扩增等位基因3～10个,平均6.1个;各位点的观察杂合度为0.057～0.574,平均为0.323;各位点PIC值在0.3723～0.8795之间,平均为0.6890。18对EST-SSR引物在长山核桃、黑核桃组、核桃楸组、铁核桃和核桃样品中,EST-SSR位点平均有效扩增率分别为87.5%、96.3%、98.8%、100%和99.8%,说明核桃EST-SSR在种、属间具有良好的通用性。聚类分析表明:在属间和核桃属内组间的聚类清晰,结果与传统分类一致。在核桃种内,参试样品可分为新疆、华北和西藏3个地理生态型,来自陕西的核桃样品与来自北京、山东、河南的样品混聚在一起,表明陕西核桃与华北核桃应属于同一地理生态型。在铁核桃种内,’金沙江铁核桃1’、’金沙江铁核桃2’与’永平铁核桃1’聚为铁核桃种。’三台’和’丽江铁核桃1’铁核桃样品,还有’丽江核桃1’与西藏核桃聚在一起,说明’丽江核桃1’应属于西藏核桃地理生态型,并且部分铁核桃与西藏核桃的亲缘关系较近。这可能是由于铁核桃与核桃混生,自然杂交使一些铁核桃后代含有了核桃基因。
Sixteen samples of Sect. Rhysocayon, 14 samples of Sect. Cardiocaryon, 28 samples of Sect. Juglans were obtained from 18 EST-SSR primers. Samples and four samples of C.illinoinensis Koch. From Carya Nutt were PCR-amplified. A total of 110 ESTs were obtained from 18 EST-SSR loci. The number of alleles per locus ranged from 3 to 10 with an average of 6.1. The observed heterozygosity at each loci ranged from 0.057 to 0.574 with an average of 0.323. The PIC value of each locus ranged from 0.3723 to 0.8795 with an average of 0.6890.18. The average effective amplification of EST-SSR primers in Changshan walnut, black walnut, walnut, walnut and walnut samples at EST-SSR loci The rates of EST-SSR of walnut EST-SSR were 87.5%, 96.3%, 98.8%, 100% and 99.8%, respectively. Cluster analysis showed that there was a clear clustering between the genus and the genus Walnut and the results were consistent with the traditional classification. In the walnut species, the test samples can be divided into three geographical ecotypes in Xinjiang, North China and Tibet. Samples of walnuts from Shaanxi and those from Beijing, Shandong and Henan were mixed together, indicating that Shaanxi walnut and North China walnut should belong to The same geographical type. Within the iron walnut species, ’Jinshajiang iron walnut 1’, ’Jinshajiang iron walnut 2’ and ’Yongping iron walnut 1’ are gathered in the walnut species. ’Santai’ and ’Lijiang iron walnut 1’ iron walnut samples, and ’Lijiang walnut 1’ together with the Tibetan walnut, ’Lijiang walnut 1’ should belong to the geographical ecology of walnut in Tibet, and some iron walnut and walnut Tibet The close relationship. This may be due to the mixture of walnuts and walnuts, and natural crosses make some of the walnuts contain walnut genes.