葡萄无核性状分子标记通用性验证

Validation of the generality of molecular markers for seedless fruit in grape

【目的】利用葡萄自然群体对已有9个无核分子标记进行评价,验证其无核检测通用性效果,为加快无核葡萄新品种选育进程提供技术支持。【方法】以88个无核种质和120个有核种质组成的自然群体为试材,对国内外已发表的9个常用葡萄无核分子标记进行通用性验证。【结果】SCAR类型标记GSLP1-569、SCC8-1080和SCF27-2000的鉴定准确率分别为57.12%、72.20%和75.38%,无核检测率分别为90.51%、79.18%和67.82%。SSR类型标记p1-VvAGL11、p2-VvAGL11、p3-VvAGL11、5U_VviAGL11、VMC7F2和VVSD10的鉴定准确率分别是88.47%、67.43%、71.94%、68.47%、67.99%和61.60%,无核检测率分别是87.74%、77.22%、90.72%、90.80%、79.10%和63.03%;卡方分析表明与无核表型极显著相关的等位点分别是250 bp、171 bp、195 bp、315 bp、197 bp和105 bp。【结论】SCAR标记SCF27-2000和SSR标记p1-VvAGL11准确率高,综合表现最优,适用于无核葡萄新品种的分子辅助选择。

【Objective】The natural population of grape(Vitis vinifera L.)was used to evaluate the universality of 9 molecular markers for seedless fruit in order to provide technical support for the breeding of new seedless grape varieties.【Methods】DNA was extracted from healthy and young samples of a natural population consisting of 88 seedless germplasmes and 120 nucleated germplasmes.PCR amplification was performed using 9 reported molecular markers for seedless fruit of grape.Then,the PCR products were detected by 1.5%agarose gel electrophoresis and capillary electrophoresis,and the specific bands were analyzed.The accuracy rate and seedless detection rate were calculated respectively to verify the versatility of 9 molecular markers for seedless fruit of grape.【Results】Among 208 grape germplasmes,16 germplasmes were detected by SCAR marker GSLP1-569,including 14 seedless germplasmes and 2 seeded germplasmes.The 14 seedless varieties were Summer Black,Changwuhebai Etc.among others.And,among them,12 germplasmes were Thompson Seedless and its derivatives.The identification accuracy and nuclear-free detection rate were 57.12%and 90.51%,respectively.Additionally,the 1080 bp specific band was amplified by SCC8-1080 in 53 seedless germplasmes and 19 seeded germplasmes,and the statistical identification accuracy and seedless detection rate were 72.20%and 79.18%,respectively.Moreover,the 2000 bp specific band was amplified by SCF27-2000 in 87 seedless germplasmes and 55 seeded germplasmes.The statistical identification accuracy and seedless detection rate were 75.38%and 67.82%,respectively.Furthermore,a total of 6 isotopic point and 8 genotypes were detected by the SSR marker p1-VvAGL11.The chi-square test showed that the allele 250 bp was significantly correlated with the nuclear-free phenotype and 257 bp was significantly correlated with nucleated phenotype.The genotype 250/250 was significantly correlated with the nuclear-free phenotype,and the genotype 257/257 was significantly correlated with the nuclear phenotype.The statistical identification accuracy and nuclear-free detection rate of the marker were 88.47%and 87.74%,respectively.A total of 3 isotopic point and 5 genotypes were detected by the marker p2-VvAGL11.The chi-square test showed that the allele 171 bp was significantly correlated with the nuclear-free phenotype.The 158 bp was significantly correlated with the nuclear phenotype.The genotype 158/171 was significantly correlated with the seedless phenotype,and the genotype 158/158 was significantly correlated with the nuclear phenotype.The accuracy of marker identification and seedless detection rate were 67.43%and 77.22%,respectively.A total of 14 isotopic point and 30 genotypes were detected by the marker p3-VvAGL11.The chi-square test showed that the allele 195 bp was significantly correlated with the nuclear-free phenotype and 185 bp was significantly correlated with the nucleated phenotype.The genotype 185/195 was significantly associated with the nuclear-free phenotype.The genotype 185/185 was significantly correlated with the nuclear phenotype,and the accuracy and nuclear-free detection rate of the marker were 71.94%and 90.72%,respectively.A total of 24 isotopic point and 75 genotypes were detected by the marker 5U_VviAGL11.The chi-square test indicated that the allele 315 bp was significantly correlated with the nuclear-free phenotype and 305 bp was significantly associated with nucleated phenotype.The genotype 307/315 was significantly correlated with the nuclear-free phenotype.The accuracy and nuclear-free detection rate of the marker were 68.47%and 90.80%,respectively.A total of 8 isotopic point were detected in the marker VMC7F2,and the allele 197 bp was significantly correlated with the seedless phenotype by chi-square test.199 bp was significantly correlated with one phenotype.The genotype 197/199 was significantly associated with the nuclear-free phenotype.The genotype 199/199 was significantly correlated with the nuclear phenotype,and the accuracy and non-nuclear detection rate of the marker were 67.99%and 79.10%,respectively.A total of 9 isotopic point and 21 genotypes were detected by the marker VVSD10.The chi-square test showed that the allele 105 bp was significantly correlated with the nuclear-free phenotype,and the genotype 105/105 was significantly correlated with the nuclear-free phenotype.The identification accuracy and nuclear-free detection rate of the marker were 61.60%and 63.03%,respectively.【Conclusion】Among the SCAR type markers,SCF27-2000 had the highest accuracy and true positive rate,and the performance was the best.And,GSLP1-569 was more suitable for the hybrid offspring of Thompson seedless series.Among the SSR markers,p1-VvAGL11 had good accuracy and seedless detection rate,and the false negative and false positive were low,showing the best performance,while p3-VvAGL11 and 5U_VviAGL11 had higher seedless detection rate,and 5U_VviAGL11 contained more genetic information.