分类： 生物学 >> 植物学 分类： 生物学 >> 植物学 分类： 农、林、牧、渔 >> 园艺学 提交时间： 2020-03-03
摘要： Background: Aneuploidy, a condition caused by an imbalance between the relative dosages of chromosomes, generally produces a novel phenotype specific to the molecular karyotype. Few techniques are currently available for detecting the molecular karyotypes of aneuploids in plants. Methods: By PCR amplification of 209 pairs of SSR primers, 17 pairs of SSR primers with no polymorphism and stable amplification in 23 different ploidy loquat were screened out. Seventeen SSR markers combined with qPCR for detection of aneuploid loquat H39 (2n=2x+5=39) karyotype variation. To verify the method for detection of aneuploid karyotype variation, 9 hybrid offspring of Q24 × ‘Huabai No. 1’ and 16 open-pollination progeny of the triploid loquat A313 and A322 were obtained, and we detected their chromosome numbers using conventional cytological methods. The SSR-qPCR method was used to detect the molecular karyotypes of aneuploids in the 9 hybrid offspring of Q24 × ‘Huabai No. 1’ and 16 open-pollination progeny of the triploid loquat A313 and A322. Results: Based on this imbalance in chromosome dosage, a new approach (referred to as ‘SSR-qPCR’) combining simple sequence repeat (SSR) markers and quantitative real-time PCR (qPCR) has been developed and utilized to detect some common aneuploids irrespective of heterozygosity. We screened 17 specific SSR markers covering all loquat linkage groups and redesigned 6 pairs of primers for SSR markers that can detect loquat chromosome aneuploidies. The SSR-qPCR detection results obtained for hybrid progeny and open-pollination progeny of triploid loquat showed diagnostic accuracies of 88.9% and 62.5%, respectively, compared with the chromosome preparation results. Limitations: Loquat genome sequences have not been published, available SSR markers are currently limited. Conclusion: SSR-qPCR can detect loquat aneuploids and be used to construct the entire molecular karyotypes of aneuploid individuals. Therefore, this method offers a novel alternative for the detection of chromosome aneuploidies.
分类： 生物学 >> 生物物理学 提交时间： 2016-05-11
Jiang, Pengfei
Zhou, Na
Chen, Xinyu
Zhao, Xing
Li, Dengyun
Wang, Fen
Zhang, Deli
Jiang, Pengfei
Bi, Lijun
摘要： H1N1 swine influenza A virus (H1N1 SwIV) is one key subtype of influenza viruses with pandemic potential. MicroRNAs (miRNAs) are endogenous small RNA molecules that regulate gene expression. MiRNAs relevant with H1N1 SwIV have rarely been reported. To understand the biological functions of miRNAs during H1N1 SwIV infection, this study profiled differentially expressed (DE) miRNAs in pulmonary alveolar macrophages from piglets during the H1N1 SwIV infection using a deep sequencing approach, which was validated by quantitative real-time PCR. Compared to control group, 70 and 16 DE miRNAs were respectively identified on post-infection day (PID) 4 and PID 7. 56 DE miRNAs were identified between PID 4 and PID 7. Our results suggest that most host miRNAs are down-regulated to defend the H1N1 SwIV infection during the acute phase of swine influenza whereas their expression levels gradually return to normal during the recovery phase to avoid the occurrence of too severe porcine lung damage. In addition, targets of DE miRNAs were also obtained, for which bioinformatics analyses were performed. Our results would be useful for investigating the functions and regulatory mechanisms of miRNAs in human influenza because pig serves as an excellent animal model to study the pathogenesis of human influenza.
点击量
待评议
点击量
下载量
评论
