Subjects: Biology >> Botany >> Applied botany submitted time 2022-04-29 Cooperative journals: 《广西植物》
Abstract： In order to explore the difference in the molecular mechanism of the variety Puerarial between isoflavones metabolic enzyme genes PtCHI, to preliminarily reveal the difference content of the isoflavones. The material of this study is Pueraria montana and Pueraria thomsonii. Puerarin and total flavonoids of Pueraria montana and Pueraria thomsonii were extracted by ethanol, and their content was measured by high-performance liquid phase. Based on the CDS sequence of the CHI gene of Pueraria montana, the PtCHI gene from Pueraria thomsonii was isolated by homologous cloning, and the protein was expressed in vitro. At the same time, the location of the PtCHI gene was studied in Arabidopsis protoplasts. The results were as follows: (1)The content of puerarin in Pueraria montana is significantly higher than the Pueraria thomsonii, and the content of total flavonoid was also, but not significant. (2)Successfully isolated the gene PtCHI from Pueraria thomsonii. The gene was 742 bp in length, contains a complete ORF frame of 672 bp, encodes 223 amino acids, and has up to 99% homology with Pueraria montana; it was predicted to be a stable hydrophobic protein with a size of 27.8 kD. (3)This study found that the expression of CHI gene in Pueraria thomsonii was stem> root> leaves, Pueraria thomsonii was root> stem> leaves. The expression of CHI genes from Pueraria thomsonii was significantly higher than Pueraria thomsonii besides in leaves; (4) Through the online tool prediction analysis, PtCHI was found to be stable hydrophilic protein and the size was 27.8 kD. The secondary and tertiary structures were based on α-Helix-based, with 25 phosphorylation sites, closely related Pueraria montana, Glycine max and Glycyrrhiza uralensis, and were more likely to interact with F3H2, F3H, 4CL4, DFR2 and CHS.(5)At the same time, the protein of PtCHI was successfully induced and isolated in vitro, with a single protein of 27.8 kD, respectively. (6) Through the Arabidopsis protoplasts revealed that PtCHI was only located in the chloroplasts. This study is in further analyzing the difference in flavonoids in Pueraria montana and Pueraria thomsonii, as well as laying the foundation for the functional verification of Pueraria thomsonii PtCHI and the research on the mechanism of isoflavone metabolism.
Subjects: Biology >> Botany >> Applied botany submitted time 2019-03-14 Cooperative journals: 《广西植物》
Abstract：以广西特有的野生柿种质‘油柿’（YS）、‘农家栽培柿’（ZP）和主栽品种‘恭城月柿’为实验材料，分析人工脱涩乙烯利处理的果实采后贮藏中可滴定酸含量、可溶性糖含量、硬度、乙烯生物合成量、色差、可溶性单宁含量、果胶及细胞壁降解酶活性的变化，探讨不同野生柿种质采后果实软化机制。研究结果表明：与恭城月柿相比，油柿可滴定酸含量高，可溶性糖含量低，转色缓慢，果实易软化。其中YS-4最不耐贮藏，乙烯利处理后，贮藏第4天硬度降至1.620 N，第6天可溶性单宁含量下降至2.398 mg·g-1，整个贮藏期原果胶含量下降最慢，末期可溶性果胶值最低，为0.832%；而YS-2最耐贮藏，乙烯利处理第8天硬度为YS-4的3.6倍，贮藏末期硬度在所有油柿中最高。农家栽培柿中乙烯利处理贮藏末期ZP-2可滴定酸含量最高，可溶性糖含量低，转色最缓慢，原果胶降幅最小，可溶性果胶含量最高；相反ZP-3果实转色快且易软化，可滴定酸含量最低，可溶性糖含量低，原果胶降幅最大，可溶性果胶含量低。恭城月柿PG活性和Cx活性远远高于油柿，各油柿种质的果实软化程度与细胞壁降解酶活性的相关性不同，β-D-Gal酶和Cx酶在农家栽培柿贮藏前期起作用，PG酶在贮藏前期和后期对果实软化起作用。说明不同柿种质品种在软化过程中起主导作用的酶不同。综上，与恭城月柿相比，‘油柿’柿耐贮性差，其中YS-4极不耐贮藏，而‘农家栽培柿’ZP-2极耐贮藏。以上结果可为研究柿果实软化机制提供基础的种质材料。
Hits
Hits
Downloads
Comment