Identification of molecular markers linked to natural astringency loss of persimmon

与柿子天然涩味丧失相关的分子标记的鉴定

• 批准号: 09556008
• 负责人: YONEMORI Keizo
• 金额: $ 9.22万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-09556008/
• 关键词: Japanese persimmon; natural astringency loss; molecular marker; PCNA; RFLP; AFLP; 日渋性マーカー; 脱渋性; ゲノム構成; 甘渋判定マーカー; Japanese persimmon; natural astringency loss; molecular marker; PCNA; RFLP; AFLP; 日渋性マーカー; 脱渋性; ゲノム構成; 甘渋判定マーカー

The pollination-constant and non-astringent (PCNA)-type fruit is the most desirable persimmon for fresh consumption, and new PCNA-type cultivars with good fruit quality have been awaited worldwide. However, the inheritance of PCNA-type is qualitative and the PCNA-type is recessive to non-PCNA (PVNA, PVA, and PCA) types. Molecular marker closely linked to the trait of natural astringency-loss is a powerful tool for the breeding project. So, we tried to establish it by the following experiments to make the breeding process more efficient.1. Bulked segregant analysis (BSA) of 6 PCNA-type and 10 non-PCNA-type offspring in breeding population was performed by AFLP method using 128 primer combinations. As a result, 4 potential AFLP markers were detected for discriminating PCNA- and non-PCNA types.2. The most reliable marker among 4 potential AFLP markers was selected and sequenced. When this marker was used as a probe for Southern analysis after genomic DNA was digested HindlII, all progenies examined could be distinguished between PCNA-type and non-PCNA type by the existence or absence of 6.5kb or 8.0 kb band. Furthermore, segregation rate of these bands indicated that persimmon shows polysomic inheritance. Persimmon seemed to be an auto- or autoallo-hexaploid.3. When Southern analysis using AFLP maker as a probe was examined to existing cultivars, PCNA cultivars could be distinguished from non-PCNA culitvars with 100% accuracy. This result indicates a high reliability of this analysis for selecting PCNA-type among wider breeding populations.4. The sequencing analysis of two RFLP markers (6.5 kb and 8.0 kb) for characterizing them and making primers for PCR analysis for easier detection of these makers was not completely successful. We could sequence 8.0 kb band but not 6.5 kb band. So, further study was needed to design PCR primers for more easier method than Southern analysis to distinguish PCNA-type from non-PCNA-type.

授粉 - 恒定和非群体（PCNA）型水果是新鲜食用的最理想的柿子，并且在全球范围内等待具有良好水果质量的新PCNA型品种。但是，PCNA型的遗传是定性的，PCNA型对非PCNA（PVNA，PVA和PCA）类型是隐性的。分子标记与自然涩味损坏的特征紧密相关的分子标记是育种项目的强大工具。因此，我们试图通过以下实验来确定它，以提高育种过程。1。使用128个引物组合，通过AFLP方法对6个PCNA型和10个非PCNA型后代进行了批量的分析分析（BSA）。结果，检测到4个潜在的AFLP标记，用于区分PCNA和非PCNA类型2。选择并测序了4个潜在AFLP标记中最可靠的标记。当对基因组DNA被消化后，将此标记用作南部分析的探针时，可以通过存在或没有6.5kb或8.0 kb频段来区分所有检查的后代。此外，这些频段的隔离率表明柿子显示了多个遗传。柿子似乎是自动或自动甲状腺素3。当将使用AFLP制造商作为探针的南部分析被检查到现有品种时，可以将PCNA品种与非PCNA Culitvars区分开，其精度为100％。该结果表明，该分析对于在更广泛的育种种群中选择PCNA型的可靠性很高。4。对两个RFLP标记（6.5 kb和8.0 kb）的测序分析，用于表征它们并进行PCR分析的引物，以便于检测这些制造商并不完全成功。我们可以对8.0 kb频段进行序列，但不能序列6.5 kb。因此，需要进一步的研究来设计PCR引物，从而比南方分析更容易地将PCNA类型与非PCNA类型区分开。