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利用SCoT、ISSR和RAPD-PCR标记对五种伽蓝菜基因型进行遗传和形态多样性评估

Genetic and Morphological Diversity Assessment of Five Kalanchoe Genotypes by SCoT, ISSR and RAPD-PCR Markers.

作者信息

Al-Khayri Jameel M, Mahdy Ehab M B, Taha Heba S A, Eldomiaty Ahmed S, Abd-Elfattah Mohamed A, Abdel Latef Arafat Abdel Hamed, Rezk Adel A, Shehata Wael F, Almaghasla Mustafa I, Shalaby Tarek A, Sattar Muhammad N, Ghazzawy Hesham S, Awad Mohamed F, Alali Khalid M, Jain Shri Mohan, Hassanin Abdallah A

机构信息

Department of Agricultural Biotechnology, College of Agriculture and Food Sciences, King Faisal University, Al-Ahsa 31982, Saudi Arabia.

National Gene Bank (NGB), Agricultural Research Centre (ARC), Giza 12613, Egypt.

出版信息

Plants (Basel). 2022 Jun 29;11(13):1722. doi: 10.3390/plants11131722.

Abstract

Determining the appropriate parents for breeding programs is the most important decision that plant breeders must make to maximize the genetic variability and produce excellent recombinant genotypes. Several methods are used to identify genotypes with desirable phenotypic features for breeding experiments. In this study, five kalanchoe genotypes were morphologically characterized by assessing plant height, number of inflorescences, number of flowers, flower length, flower diameter and number of petals. The analysis showed the distinction of yellow kalanchoe in the plant height trait, while the orange kalanchoe was distinguished in the number of inflorescences, the number of flowers and flower length traits, whereas the violet kalanchoe possessed the largest flower diameter and the highest number of petals. The molecular profiling was performed by random amplified polymorphism DNA (RAPD), inter-simple sequence repeats (ISSR) and start codon targeted (SCoT)-polymerase chain reaction (PCR) tools. Genomic DNA was extracted from young leaves and the PCR reactions were performed using ten primers for each SCoT, ISSR and RAPD marker. Only four out of ten primers showed amplicon profiles in all PCR markers. A total of 70 bands were generated by SCoT, ISSR and RAPD-PCR with 35 polymorphic bands and 35 monomorphic bands. The total number of bands of RAPD, ISSR and SCoT was 15, 17 and 38, respectively. The polymorphism percentages achieved by RAPD, ISSR and SCoT were 60.25%, 15% and 57%, respectively. The cluster analysis based on morphological data revealed two clusters. Cluster I consisted of violet and orange kalanchoe, and cluster II comprised red, yellow and purple kalanchoe. Whereas the cluster analysis based on molecular data revealed three clusters. Cluster I included only yellow kalanchoe, cluster II comprised orange and violet kalanchoe while cluster III comprised red, and purple kalanchoe. The study concluded that orange, violet and yellow kalanchoe are distinguished parents for breeding economically valued traits in kalanchoe. Also, the study concluded that SCoT and RAPD markers reproduced reliable banding patterns to assess the genetic polymorphism among kalanchoe genotypes that consider the basis stone for genetic improvements in ornamental plants.

摘要

确定育种计划的合适亲本是植物育种者为使遗传变异性最大化并培育出优良重组基因型而必须做出的最重要决定。有几种方法可用于识别具有理想表型特征的基因型以进行育种实验。在本研究中,通过评估株高、花序数、花数、花长、花径和花瓣数对5种长寿花基因型进行了形态学特征分析。分析表明,黄色长寿花在株高性状上有区别,橙色长寿花在花序数、花数和花长性状上有区别,而紫色长寿花的花径最大且花瓣数最多。通过随机扩增多态性DNA(RAPD)、简单序列重复区间(ISSR)和起始密码子靶向(SCoT)-聚合酶链反应(PCR)工具进行分子图谱分析。从幼叶中提取基因组DNA,并使用针对每个SCoT、ISSR和RAPD标记的10种引物进行PCR反应。10种引物中只有4种在所有PCR标记中显示出扩增子图谱。SCoT、ISSR和RAPD-PCR共产生70条带,其中35条多态性带和35条单态性带。RAPD、ISSR和SCoT的条带总数分别为15条、17条和38条。RAPD、ISSR和SCoT的多态性百分比分别为60.25%、15%和57%。基于形态学数据的聚类分析显示有两个聚类。聚类I由紫色和橙色长寿花组成,聚类II由红色、黄色和紫色长寿花组成。而基于分子数据的聚类分析显示有三个聚类。聚类I仅包括黄色长寿花,聚类II由橙色和紫色长寿花组成,聚类III由红色和紫色长寿花组成。该研究得出结论,橙色、紫色和黄色长寿花是培育长寿花经济价值性状的优良亲本。此外,该研究得出结论,SCoT和RAPD标记产生了可靠的条带模式,以评估长寿花基因型之间的遗传多态性,这是观赏植物遗传改良的基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8139/9268761/61722bcc3406/plants-11-01722-g001.jpg

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