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特有物种舒尔特(Schult.)自然种群遗传多样性的引物结合位点(PBS)分析

Primer Binding Site (PBS) Profiling of Genetic Diversity of Natural Populations of Endemic Species Schult.

作者信息

Khapilina Oxana, Turzhanova Ainur, Danilova Alevtina, Tumenbayeva Asem, Shevtsov Vladislav, Kotukhov Yuri, Kalendar Ruslan

机构信息

National Center for Biotechnology, Korgalzhin Hwy 13/5, Nur-Sultan 010000, Kazakhstan.

Altai Botanical Garden, Yermakova Str 1, Ridder 070000, Kazakhstan.

出版信息

BioTech (Basel). 2021 Oct 13;10(4):23. doi: 10.3390/biotech10040023.

Abstract

Endemic species are especially vulnerable to biodiversity loss caused by isolation or habitat specificity, small population size, and anthropogenic factors. Endemic species biodiversity analysis has a critically important global value for the development of conservation strategies. The rare onion is a narrow-lined endemic species, with natural populations located in the extreme climatic conditions of the Kazakh Altai. populations are decreasing everywhere due to anthropogenic impact, and therefore, this species requires preservation and protection. Conservation of this rare species is associated with monitoring studies to investigate the genetic diversity of natural populations. Fundamental components of eukaryote genome include multiple classes of interspersed repeats. Various PCR-based DNA fingerprinting methods are used to detect chromosomal changes related to recombination processes of these interspersed elements. These methods are based on interspersed repeat sequences and are an effective approach for assessing the biological diversity of plants and their variability. We applied DNA profiling approaches based on conservative sequences of interspersed repeats to assess the genetic diversity of natural populations located in the territory of Kazakhstan Altai. The analysis of natural populations, carried out using the DNA profiling approach, allowed the effective differentiation of the populations and assessment of their genetic diversity. We used conservative sequences of tRNA primer binding sites (PBS) of the long-terminal repeat (LTR) retrotransposons as PCR primers. Amplification using the three most effective PBS primers generated 628 PCR amplicons, with an average of 209 amplicons. The average polymorphism level varied from 34% to 40% for all studied samples. Resolution analysis of the PBS primers showed all of them to have high or medium polymorphism levels, which varied from 0.763 to 0.965. Results of the molecular analysis of variance showed that the general biodiversity of populations is due to interpopulation (67%) and intrapopulation (33%) differences. The revealed genetic diversity was higher in the most distant population of LD64, located on the Sarymsakty ridge of Southern Altai. This is the first genetic diversity study of the endemic species using DNA profiling approaches. This work allowed us to collect new genetic data on the structure of populations in the Altai for subsequent development of preservation strategies to enhance the reproduction of this relict species. The results will be useful for the conservation and exploitation of this species, serving as the basis for further studies of its evolution and ecology.

摘要

特有物种特别容易受到因隔离或栖息地特异性、种群规模小以及人为因素导致的生物多样性丧失的影响。特有物种生物多样性分析对于制定保护策略具有至关重要的全球价值。珍稀洋葱是一种分布狭窄的特有物种,其天然种群位于哈萨克阿尔泰山脉的极端气候条件下。由于人为影响,各地的种群数量都在减少,因此,该物种需要得到保护。对这种珍稀物种的保护与监测研究相关,以调查天然种群的遗传多样性。真核生物基因组的基本组成部分包括多类散布重复序列。各种基于聚合酶链反应(PCR)的DNA指纹图谱方法被用于检测与这些散布元件重组过程相关的染色体变化。这些方法基于散布重复序列,是评估植物生物多样性及其变异性的有效途径。我们应用基于散布重复保守序列的DNA分析方法来评估位于哈萨克阿尔泰山地区的天然种群的遗传多样性。使用DNA分析方法对天然种群进行的分析,能够有效区分种群并评估其遗传多样性。我们使用长末端重复(LTR)逆转座子的转运RNA引物结合位点(PBS)的保守序列作为PCR引物。使用三种最有效的PBS引物进行扩增产生了628个PCR扩增产物,平均为209个扩增产物。所有研究样本的平均多态性水平在34%至40%之间。对PBS引物的分辨率分析表明,它们都具有高或中等多态性水平,范围从0.763到0.965。方差分子分析结果表明,种群的总体生物多样性归因于种群间(67%)和种群内(33%)的差异。在位于南阿尔泰山脉萨雷姆萨克蒂山脊的最偏远种群LD64中,所揭示的遗传多样性更高。这是首次使用DNA分析方法对该特有物种进行的遗传多样性研究。这项工作使我们能够收集有关阿尔泰山种群结构的新遗传数据,以便随后制定保护策略,以促进这种残遗物种的繁殖。研究结果将有助于该物种的保护和利用,为其进化和生态学的进一步研究奠定基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2c5/9245474/fb5c1599aca5/biotech-10-00023-g001.jpg

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