苦荞黄烷酮3-羟化酶基因FtF3H及其启动子的克隆与功能分析

【目的】克隆苦荞FtF3H基因及其启动子序列，探究该基因的生物学效应及启动子对环境因素与激素的响应。【方法】基于苦荞基因组和转录组数据克隆FtF3H基因及其启动子，对二者进行生物信息学分析，采用qRT-PCR技术检测FtF3H对MeJA、ABA和光处理的响应，进一步利用转基因拟南芥技术，分析FtF3H的过表达对黄酮合成的影响。【结果】苦荞FtF3H基因DNA序列长2 034 bp，包括2个内含子和3个外显子；其ORF序列为1 104 bp，推导的FtF3H蛋白含367个氨基酸残基，归类于2-酮戊二酸依赖性双加氧酶超家族且具有典型的植物F3H的分子特征。启动子PFtF3H在核心启动子基序外，还含有数目众多的光应答元件和激素响应元件，qRT-PCR检测结果显示，PFtF3H可强烈响应ABA、MeJA和光照的诱导。转基因拟南芥中，FtF3H的过表达可引起总黄酮含量显著增加（P<0.05），黄酮合成相关酶基因，AtANS、AtDFR和AtF′3H等的表达量显著上调（P<0.05），而拟南芥内源的AtF3H表达受到抑制（P<0.05）。【结论】苦荞FtF3H基因是一个典型的植物...     

Effects of dietary concentrate level and chromium-methionine supplementation on ruminal fermentation,and ruminal bacterial and fatty acid composition in Tan lambs北大核心CSCD

This study evaluated the effects of dietary concentrate level and chromium methionine(Cr-Met)supplementation on ruminal fermentation and fatty acid composition, and ruminal bacteria abundance in Tan lambs. Forty male Tan lambs [(21.00±1.23) kg body weight, 5 months of age] were randomly assigned to four treatment groups with 10 lambs in each treatment. The treatments were: a low-concentrate (LC) diet (concentrate:forage, 35:65) without Cr-Met supplementation; a high-concentrate (HC) diet (concentrate:forage, 55:45) without Cr-Met supplementation; groups fed the HC diet with 0.75 or 1.50 g·d-1·lamb-1 Cr-Met (HCM and HCH, respectively). Ruminal fluid was collected on day 65 with an oral stomach tube about 3 h after the morning feed and samples used for fermentation analysis, fatty acid composition determination and bacterial DNA extraction. It was found that: 1) Ruminal pH, acetate proportion and acetate:propionate were greater (P<0.05) with the LC diet whereas microbial crude protein, propionate and valerate proportions were greater (P<0.05) with the HC diet. The acetate:propionate was greater (P<0.05) in the HCM group than in the HC group. 2) For the HC diet, the DNA abundances of Butyrivibrio fibrisolvens vaccenic acid subgroup (Butyrivibrio VA) and Ruminococcus flavefaciens were decreased (P<0.05), compared with the LC diet, whereas the DNA abundance of Anaerovibrio lipolytica was increased (P<0.05). For lambs fed the HC diet, DNA abundances of B. fibrisolvens stearic acid subgroup, Butyrivibrio VA, Butyrivibrio proteoclasticus, and A. lipolytica showed a linear decrease (P<0.05) with increasing Cr-Met supplementation level; However, the DNA abundance of R. flavefaciens showed a linear increase (P<0.05). 3) Compared with the LC group, the concentrations of t11 C18:1, trans C18:1, c9t11 CLA, t10c12 CLA, C18:2n6, and C18:3n3 were decreased by the HC diet without Cr-Met supplementation whereas the concentration of C18:0 was increased. There was a linear increase in t11 C18:1 and trans C18:1 with increasing Cr-Met supplementation level in lambs fed the HC diet. The results suggest that the HC diet inhibited the growth of ruminal bacteria involved in the ruminal biohydrogenation process, and the addition of Cr-Met had a potentially positive effect on conjugated linoleic acid (CLA) synthesis in body tissue. © 2022, Editorial Office of Acta Prataculturae Sinica. All rights reserved.