[Objective] This study aimed to clone a full-length CHS gene from buck- wheat. [Method] With total RNA extracted from buckwheat as the template, CHS cD- NA sequence was cloned from buckwheat by using RACE technology a...[Objective] This study aimed to clone a full-length CHS gene from buck- wheat. [Method] With total RNA extracted from buckwheat as the template, CHS cD- NA sequence was cloned from buckwheat by using RACE technology and CODEHOP primer design method, the full length gene was obtained by primers which were de- signed for amplification of full-length gene sequence with buckwheat DNA template. Clustalxl.81 and MEGA4 software were used for sequence analysis and construction of phylogenetic tree; NCBI Blastn and Biastp programs were applied for homology analysis of nucleic acid and protein. [Result] Bioinformatics analysis showed that the full length of this gene is 1 906 bp, containing a 463 bp intron sequence and a 1 188 bp coding region, encoding 395 amino acids. Blastn sequence alignment revealed that the CHS gene sequence obtained in this study shared 86% homology with the CHS gene of closely related species. [Conclusion] This study laid the foundation to clarify molecular basis of the synthesis of buckwheat bioflavanoids and explore an effective way to improve the content of buckwheat bioflavanoids.展开更多
Heterotrichous ciliates play an important role in aquatic ecosystem energy flow processes and many are model organisms for research in cytology,regenerative biology,and toxicology.In the present study,we combine both ...Heterotrichous ciliates play an important role in aquatic ecosystem energy flow processes and many are model organisms for research in cytology,regenerative biology,and toxicology.In the present study,we combine both morphological and molecular data to infer phylogenetic relationships at family-genus level and propose new evolutionary hypotheses for the class Heterotrichea.The main results include:(1)96 new ribosomal DNA sequences from 36 populations,representing eight families and 13 genera,including three poorly annotated genera,Folliculinopsis,Ampullofolliculina and Linostomella;(2)the earliest-branching families are Spirostomidae in single-gene trees and Peritromidae in the concatenated tree,but the family Peritromidae probably represents the basal lineage based on its possession of many"primitive"morphological characters;(3)some findings in molecular trees are not supported by morphological evidence,such as the family Blepharismidae is one of the most recent branches and the relationship between Fabreidae and Folliculinidae is very close;(4)the systematic positions of Condylostomatidae,Climacostomidae,and Gruberiidae remain uncertain based either on morphological or molecular data;and(5)the monophyly of each genus included in the present study is supported by the molecular phylogenetic trees,except for Blepharisma in the SSU r DNA tree and Folliculina in the ITS1-5.8 S-ITS2 tree.展开更多
基金Supported by 948 Program,Ministry of Agriculture of China(2008-z27)~~
文摘[Objective] This study aimed to clone a full-length CHS gene from buck- wheat. [Method] With total RNA extracted from buckwheat as the template, CHS cD- NA sequence was cloned from buckwheat by using RACE technology and CODEHOP primer design method, the full length gene was obtained by primers which were de- signed for amplification of full-length gene sequence with buckwheat DNA template. Clustalxl.81 and MEGA4 software were used for sequence analysis and construction of phylogenetic tree; NCBI Blastn and Biastp programs were applied for homology analysis of nucleic acid and protein. [Result] Bioinformatics analysis showed that the full length of this gene is 1 906 bp, containing a 463 bp intron sequence and a 1 188 bp coding region, encoding 395 amino acids. Blastn sequence alignment revealed that the CHS gene sequence obtained in this study shared 86% homology with the CHS gene of closely related species. [Conclusion] This study laid the foundation to clarify molecular basis of the synthesis of buckwheat bioflavanoids and explore an effective way to improve the content of buckwheat bioflavanoids.
基金supported by the Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology(Qingdao)(2018SDKJ0406-1)the National Natural Science Foundation of China(3203000059,31970398)。
文摘Heterotrichous ciliates play an important role in aquatic ecosystem energy flow processes and many are model organisms for research in cytology,regenerative biology,and toxicology.In the present study,we combine both morphological and molecular data to infer phylogenetic relationships at family-genus level and propose new evolutionary hypotheses for the class Heterotrichea.The main results include:(1)96 new ribosomal DNA sequences from 36 populations,representing eight families and 13 genera,including three poorly annotated genera,Folliculinopsis,Ampullofolliculina and Linostomella;(2)the earliest-branching families are Spirostomidae in single-gene trees and Peritromidae in the concatenated tree,but the family Peritromidae probably represents the basal lineage based on its possession of many"primitive"morphological characters;(3)some findings in molecular trees are not supported by morphological evidence,such as the family Blepharismidae is one of the most recent branches and the relationship between Fabreidae and Folliculinidae is very close;(4)the systematic positions of Condylostomatidae,Climacostomidae,and Gruberiidae remain uncertain based either on morphological or molecular data;and(5)the monophyly of each genus included in the present study is supported by the molecular phylogenetic trees,except for Blepharisma in the SSU r DNA tree and Folliculina in the ITS1-5.8 S-ITS2 tree.
