中国甜柿自然脱涩性状早期筛选及其杂交育种研究
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摘要
栽培柿(Diospyros kaki Thunb.)品种多,变异丰富,为杂交育种研究提供了丰富的亲本材料。但其倍性高(6x)、雄花资源少,也是其遗传改良的主要障碍。本研究在对中国甜柿育种亲本组合进行分析的基础上,重点对其自然脱涩性状的早期选择进行了研究；此外,利用SCoT技术对部分柿品种的鉴定和遗传关系进行了探讨；以期为中国甜柿遗传改良提供科学依据。主要结果如下：
1.应用36条引物对31个柿品种和3个柿近缘种进行SCoT分析。从36条引物中筛选得到17条有多态性、重复性好的引物；34份试材共扩增出154条谱带,平均多态性比率94.16%,每引物可产生8.5条多态性谱带；2种聚类方法均表明来自江西省的种质独自聚为一大组,且组内又按种质的地理分布各自聚类；完全甜柿与非完全甜柿截然分开,且中国甜柿与日本甜柿亦各自聚类。
2.统计8份杂交父本的花粉离体萌发率并计算所有杂交组合的坐果率、得胚数及亲和指数,发现供试8份雄性种质花粉离体萌发率在16.8%-64.2%之间；中国原产完全雄株种质花粉的离体萌发率与常见授粉品种‘禅寺丸’和‘西村早生’相当；同一种质在不同年份的花粉离体萌发率差异不明显。坐果率受母本影响更大；坐果率与亲和指数之间无明显相关性；品种的单性结实能力对亲和指数影响较大。
3.对不同脱涩类型的品种进行Mast/MAST及R02位点基因分型,发现Mast/MAST不能用于鉴定中国甜柿的脱涩类型；供试44份柿品种(包括甜涩性状已知的所有中国甜柿类型)、2份柿属植物(君迁子和油柿)和两个杂交F1群体(共117株)中,10份中国甜柿和6份完全雄性种质可检测到R02标记,说明该标记可能与控制中国甜柿自然脱涩性状相关等位基因连锁；R02标记在两个杂交F1群体均呈现1：1比例分离,说明中国甜柿自然脱涩性状受单个显性基因位点控制；完全雄性种质亦可检测到自然脱涩基因标记,可能说明其作为完全甜柿育种亲本的应用潜力。
综上所述,进行中国甜柿杂交育种工作最有前景的的杂交组合是‘鄂柿1号’、‘恭城水柿’、‘华柿1号’分别与‘罗田甜柿’和含有R02标记的完全雄性种质的组合；中国甜柿快速遗传改良体系与日本柿品种选育程序相比,不仅克服了近亲退化的现象,还缩短了育种时问,提高了育种效率。
There is abundant persimmon(Diospyros kaki Thunb.) cultivars existing in China and natural variation provide plenty of material for cross-breeding research. There are two main obstacles in persimmon genetic improvement, one is the high ploidy and the other one is fewer staminate germplasm. In the present study, on the basis of the analyzing the parents combination of Chinese PCNA cross breeding, we focused on the early screening of natural deastringency trait. In addition, we investigated the identification and genetic relationships among some persimmon cultivar by SCoT, hoping provided a scientific basis for Chinese PCNA genetic improvement. The main results were as follows:
1. A total of154bands were generated by17informative SCoT primers screened from36SCoT primers in34Diospyros spp. The polymorphic rate of SCoT markers was94.16%, and the average number of polymorphisms was8.5. The result of clustering analysis using UPGMA and PCoA showed that the34genotypes could be divided into3Clusters. The first cluster included the persimmon local cultivar from Jiangxi province, while the second consisted of other persimmon cultivar.. Oil persimmon, Chekiang persimmon and Jinzaoshi formed the third group. PCNA can be separated from non-PCNA, and PCNA originated from Japan and China was distinguished. Cluster analysis of SCoT markers through the UPGMA and PCoA analysis were largely consistent.
2. Eight Diospyros spp. staminate germplasm materials were used to test they pollen germinability in vitro. The results showed that the pollen germinability in vitro of those materials from16.8%to64.2%. The pollen germinability between Chinese androecious genotypes and common pollination cultivar 'Zenjimaru' and 'Nishimura-wase' were equal. There was not much difference about the pollen germinability in a same genotype at different years. Fruit setting rate was impacted greatly by maternal. There was no relevant between affinity index and fruit setting rate. The parthenocarpic ability could impact the affinity index.
3. The astringency type of Chinese PCNA persimmon can not be identified throughout Mast/MAST genotyping; the RO2marker was detected in ten Chinese PCNA types and six androecious genotypes from44persimmon cultivars, but not in any of the non-PNCA and Japanese PCNA types.This indicated that the RO2maybe linked to the allele associated with the non-astringent trait of the Chinese PCNA types. The segregation behavior of RO2in both Fi populations was consistent with a1:1ratio, suggesting the non-astringent trait of Chinese PCNA types is controlled by a single domiant gene locus. The presence of the RO2marker among androecious material indicates that they also have potential as donor of non-astringency trait of Chinese PCNA type.
In summary, the most valueable combinations of Chinese PCNA cross breeding maybe from 'Eshi No.l','Gongcheng-shuishi','Huashi No.l'crossing with'Luotian-tianshi'and Chinese androecious genotypes respectively. Compared to persimmon breeding program in NIFTS, there are two advantage in genetic improvement system of Chinese PCNA persimmon, one is avoiding inbreed depress, and the other one is improving the efficiency of PCNA breeding.
1.应用36条引物对31个柿品种和3个柿近缘种进行SCoT分析。从36条引物中筛选得到17条有多态性、重复性好的引物；34份试材共扩增出154条谱带,平均多态性比率94.16%,每引物可产生8.5条多态性谱带；2种聚类方法均表明来自江西省的种质独自聚为一大组,且组内又按种质的地理分布各自聚类；完全甜柿与非完全甜柿截然分开,且中国甜柿与日本甜柿亦各自聚类。
2.统计8份杂交父本的花粉离体萌发率并计算所有杂交组合的坐果率、得胚数及亲和指数,发现供试8份雄性种质花粉离体萌发率在16.8%-64.2%之间；中国原产完全雄株种质花粉的离体萌发率与常见授粉品种‘禅寺丸’和‘西村早生’相当；同一种质在不同年份的花粉离体萌发率差异不明显。坐果率受母本影响更大；坐果率与亲和指数之间无明显相关性；品种的单性结实能力对亲和指数影响较大。
3.对不同脱涩类型的品种进行Mast/MAST及R02位点基因分型,发现Mast/MAST不能用于鉴定中国甜柿的脱涩类型；供试44份柿品种(包括甜涩性状已知的所有中国甜柿类型)、2份柿属植物(君迁子和油柿)和两个杂交F1群体(共117株)中,10份中国甜柿和6份完全雄性种质可检测到R02标记,说明该标记可能与控制中国甜柿自然脱涩性状相关等位基因连锁；R02标记在两个杂交F1群体均呈现1：1比例分离,说明中国甜柿自然脱涩性状受单个显性基因位点控制；完全雄性种质亦可检测到自然脱涩基因标记,可能说明其作为完全甜柿育种亲本的应用潜力。
综上所述,进行中国甜柿杂交育种工作最有前景的的杂交组合是‘鄂柿1号’、‘恭城水柿’、‘华柿1号’分别与‘罗田甜柿’和含有R02标记的完全雄性种质的组合；中国甜柿快速遗传改良体系与日本柿品种选育程序相比,不仅克服了近亲退化的现象,还缩短了育种时问,提高了育种效率。
There is abundant persimmon(Diospyros kaki Thunb.) cultivars existing in China and natural variation provide plenty of material for cross-breeding research. There are two main obstacles in persimmon genetic improvement, one is the high ploidy and the other one is fewer staminate germplasm. In the present study, on the basis of the analyzing the parents combination of Chinese PCNA cross breeding, we focused on the early screening of natural deastringency trait. In addition, we investigated the identification and genetic relationships among some persimmon cultivar by SCoT, hoping provided a scientific basis for Chinese PCNA genetic improvement. The main results were as follows:
1. A total of154bands were generated by17informative SCoT primers screened from36SCoT primers in34Diospyros spp. The polymorphic rate of SCoT markers was94.16%, and the average number of polymorphisms was8.5. The result of clustering analysis using UPGMA and PCoA showed that the34genotypes could be divided into3Clusters. The first cluster included the persimmon local cultivar from Jiangxi province, while the second consisted of other persimmon cultivar.. Oil persimmon, Chekiang persimmon and Jinzaoshi formed the third group. PCNA can be separated from non-PCNA, and PCNA originated from Japan and China was distinguished. Cluster analysis of SCoT markers through the UPGMA and PCoA analysis were largely consistent.
2. Eight Diospyros spp. staminate germplasm materials were used to test they pollen germinability in vitro. The results showed that the pollen germinability in vitro of those materials from16.8%to64.2%. The pollen germinability between Chinese androecious genotypes and common pollination cultivar 'Zenjimaru' and 'Nishimura-wase' were equal. There was not much difference about the pollen germinability in a same genotype at different years. Fruit setting rate was impacted greatly by maternal. There was no relevant between affinity index and fruit setting rate. The parthenocarpic ability could impact the affinity index.
3. The astringency type of Chinese PCNA persimmon can not be identified throughout Mast/MAST genotyping; the RO2marker was detected in ten Chinese PCNA types and six androecious genotypes from44persimmon cultivars, but not in any of the non-PNCA and Japanese PCNA types.This indicated that the RO2maybe linked to the allele associated with the non-astringent trait of the Chinese PCNA types. The segregation behavior of RO2in both Fi populations was consistent with a1:1ratio, suggesting the non-astringent trait of Chinese PCNA types is controlled by a single domiant gene locus. The presence of the RO2marker among androecious material indicates that they also have potential as donor of non-astringency trait of Chinese PCNA type.
In summary, the most valueable combinations of Chinese PCNA cross breeding maybe from 'Eshi No.l','Gongcheng-shuishi','Huashi No.l'crossing with'Luotian-tianshi'and Chinese androecious genotypes respectively. Compared to persimmon breeding program in NIFTS, there are two advantage in genetic improvement system of Chinese PCNA persimmon, one is avoiding inbreed depress, and the other one is improving the efficiency of PCNA breeding.
引文
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