Nyaku Seloame T, Karapareddy Sowndarya, Cebert Ernst, Lawrence Kathy, Eleblu John S Y, Sharma Govind C, Sripathi Venkateswara R
Department of Crop Science, College of Basic and Applied Sciences, University of Ghana, Legon, Accra P.O. Box LG 44, Ghana.
Department of Biological and Environmental Sciences, Alabama A&M University, Huntsville, AL 35811, USA.
Plants (Basel). 2023 Dec 19;13(1):5. doi: 10.3390/plants13010005.
Around 300 different plant species are infected by the plant-parasitic reniform nematode (), including cotton. This is a devasting nematode with a preference for cotton; it is commonly found in Alabama farms and causes severe reduction in yields. Its first internal transcribed spacer (ITS1) region can be sequenced, and potential mutations can be found in order to study the population dynamics of the reniform nematode. The goal of our study was to sequence the ITS1 rDNA region in male and female RNs that were collected from BelleMina, Hamilton, and Lamons locations in Alabama. After separating the single male and female RNs from the samples collected from the three selected listed sites above, the ITS1 region was amplified selectively using specific primers, and the resulting products were cloned and sequenced. Two distinct bands were observed after DNA amplification of male and female nematodes at 550 bp and 730 bp, respectively. The analysis of sequenced fragments among the three populations showed variation in average nucleotide frequencies of female and male RNs. Singletons within the female and male Hamilton populations ranged from 7.8% to 10%, and the variable sites ranged from 13.4% to 26%. However, female and male BelleMina populations had singletons ranging from 7.1% to 19.7% and variable regions in the range of 13.9% to 49.3%. The female and male Lamons populations had singletons ranging from 2.5% to 8.7% and variable regions in the range of 2.9% to 14.2%. Phylogenetic (neighbor-joining) analysis for the two ITS1 fragments (ITS-550 and ITS-730) showed relatively high intra-nematode variability. Different clone sequences from an individual nematode often had greater similarity with other nematodes than with their own sequences. RNA fold analysis of the ITS1 sequences revealed varied stem and loop structures, suggesting both conserved and variable regions in the variants identified from female and male RNs, thus underscoring the presence of significant intra- and inter-nematodal variation among RN populations in Alabama.
大约300种不同的植物物种会被植物寄生肾形线虫（ ）感染，其中包括棉花。这是一种对棉花有偏好的毁灭性线虫；它在阿拉巴马州的农场很常见，会导致产量大幅下降。可以对其第一个内部转录间隔区（ITS1）进行测序，并发现潜在的突变，以便研究肾形线虫的种群动态。我们研究的目的是对从阿拉巴马州的贝勒米纳、汉密尔顿和拉蒙斯地区采集的雄性和雌性肾形线虫的ITS1 rDNA区域进行测序。从上述三个选定地点采集的样本中分离出单个雄性和雌性肾形线虫后，使用特异性引物选择性扩增ITS1区域，并对所得产物进行克隆和测序。在对雄性和雌性线虫进行DNA扩增后，分别在550 bp和730 bp处观察到两条不同的条带。对三个种群中测序片段的分析表明，雌性和雄性肾形线虫的平均核苷酸频率存在差异。汉密尔顿种群中雌性和雄性的单例范围为7.8%至10%，可变位点范围为13.4%至26%。然而，贝勒米纳种群的雌性和雄性单例范围为7.1%至19.7%，可变区域范围为13.9%至49.3%。拉蒙斯种群的雌性和雄性单例范围为2.5%至8.7%，可变区域范围为2.9%至14.2%。对两个ITS1片段（ITS - 550和ITS - 730）进行的系统发育（邻接法）分析显示，线虫内部的变异性相对较高。来自单个线虫的不同克隆序列通常与其他线虫的相似性高于与其自身序列的相似性。对ITS1序列的RNA折叠分析揭示了不同的茎环结构，表明从雌性和雄性肾形线虫中鉴定出的变体中存在保守区域和可变区域，从而强调了阿拉巴马州肾形线虫种群内部和种群之间存在显著变异。
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