本文主要研究内容
作者王平（2019）在《碳代谢阻遏基因snf1和Reg1调控黄曲霉生长发育和黄曲霉毒素合成》一文中研究指出：黄曲霉毒素具有强致癌性和强毒性,严重威胁食品安全和人畜健康。碳代谢阻遏物（CreA）是真菌碳代谢过程关键调控因子,是协调水解酶合成分泌、碳代谢和次级代谢产物合成的关键节点。AMP依赖的蛋白激酶（SNF1）和磷酸酶Ⅰ（Reg1）可调控CreA磷酸化水平,影响CreA细胞定位和功能。本研究通过构建缺失突变株,解析SNF1和Reg1对黄曲霉生长发育、碳源利用和黄曲霉毒素合成的调控作用。主要结论如下:（1）在黄曲霉野生株NRRL3357中敲除snf1基因后,突变株在马铃薯葡萄糖琼脂培养基、葡萄糖、蔗糖、海藻糖、淀粉、果胶和微晶纤维素培养基中,菌落直径普遍变小,菌落颜色变浅,分生孢子量减少。敲除Reg1基因后,在葡萄糖、蔗糖、海藻糖、果胶和微晶纤维素培养基上,菌落直径比出发菌株小,孢子量减少;在淀粉培养基上,菌落直径略微变大,孢子在菌落边缘形成完整的环状。结合显微观察实验的结果,表明SNF1和Reg1正调控黄曲霉的生长和分生孢子的正常发育。（2）淀粉培养基上,Δsnf1菌落直径减小,ΔReg1菌落直径变大,SNF1和Reg1可能通过碳代谢抑制因子CreA影响淀粉酶表达,Reg1基因敲除后导致核内CreA含量下降,诱导淀粉水解酶的表达分泌,而snf1基因敲除后导致核内CreA含量上升,抑制淀粉水解酶的表达分泌。果胶、微晶纤维素多糖培养基上,Δsnf1和ΔReg1生长速度出现差异都呈现下降趋势,SNF1和Reg1通过其他途径参与胞外水解酶表达调控。（3）对突变株Δsnf1和ΔReg1黄曲霉毒素B1（AFB1）产量分析发现,AFB1产量均下降,降低率分别为80.2%,77.4%（以产毒培养基YES为例）,表明SNF1和Reg1对毒素AFB1合成具有正调控作用。（4）分析突变株Δsnf1和ΔReg1毒素合成相关基因aflD、aflM、aflO、aflP和转录因子aaflS/flR基因和调控分生孢子转录基因brlA、abaA的转录水平可以知道:snf1、Reg1基因敲除后下调转录因子aflS/aflR的转录表达水平,进而结构基因转录受到抑制,最终引起毒素合成受阻。在aflS/aflR启动子区存在CreA结合位点,SNF1和Reg1缺失后影响CreA磷酸化水平,扰乱CreA在核质间的动态平衡,影响转录因子aflS/aflR的转录表达水平。在Δsnf1和ΔReg1突变体中,参与调控分生孢子形成发育基因呈现下调。snf1基因缺失引起分生孢子滞后发育形成,结合显微观察发现分生孢子缺陷。因此我们推断,snf1调控分生孢子的准备阶段,该基因缺失导致生长分化相关基因下调,如brlA、abaA转录抑制造成分生孢子滞后发育。（5）采用侵染花生实验,研究黄曲霉SNF1和Reg1缺失对黄曲霉侵染定殖力的影响。研究发现,snf1缺失后花生表面几乎没有黄曲霉生长,黄曲霉侵染定殖受到极大程度抑制,并且检测不到AFB1。Reg1缺失后花生少部分表面被黄曲霉包裹,生长受到抑制,AFB1含量急剧下降。表明SNF1和Reg1对于黄曲霉侵染定殖能力具有正调控作用。SNF1和Reg1影响水解酶,如淀粉酶、果胶酶等的表达分泌,最终削弱黄曲霉侵染和定殖能力,比较而言SNF1缺失对黄曲霉侵染定殖能力造成的影响更大。本研究结果为黄曲霉毒素污染风险评估及防控技术开发奠定一定的理论基础。
Abstract
huang qu mei du su ju you jiang zhi ai xing he jiang du xing ,yan chong wei xie shi pin an quan he ren chu jian kang 。tan dai xie zu e wu （CreA）shi zhen jun tan dai xie guo cheng guan jian diao kong yin zi ,shi xie diao shui jie mei ge cheng fen bi 、tan dai xie he ci ji dai xie chan wu ge cheng de guan jian jie dian 。AMPyi lai de dan bai ji mei （SNF1）he lin suan mei Ⅰ（Reg1）ke diao kong CreAlin suan hua shui ping ,ying xiang CreAxi bao ding wei he gong neng 。ben yan jiu tong guo gou jian que shi tu bian zhu ,jie xi SNF1he Reg1dui huang qu mei sheng chang fa yo 、tan yuan li yong he huang qu mei du su ge cheng de diao kong zuo yong 。zhu yao jie lun ru xia :（1）zai huang qu mei ye sheng zhu NRRL3357zhong qiao chu snf1ji yin hou ,tu bian zhu zai ma ling shu pu tao tang qiong zhi pei yang ji 、pu tao tang 、zhe tang 、hai zao tang 、dian fen 、guo jiao he wei jing qian wei su pei yang ji zhong ,jun la zhi jing pu bian bian xiao ,jun la yan se bian jian ,fen sheng bao zi liang jian shao 。qiao chu Reg1ji yin hou ,zai pu tao tang 、zhe tang 、hai zao tang 、guo jiao he wei jing qian wei su pei yang ji shang ,jun la zhi jing bi chu fa jun zhu xiao ,bao zi liang jian shao ;zai dian fen pei yang ji shang ,jun la zhi jing lve wei bian da ,bao zi zai jun la bian yuan xing cheng wan zheng de huan zhuang 。jie ge xian wei guan cha shi yan de jie guo ,biao ming SNF1he Reg1zheng diao kong huang qu mei de sheng chang he fen sheng bao zi de zheng chang fa yo 。（2）dian fen pei yang ji shang ,Δsnf1jun la zhi jing jian xiao ,ΔReg1jun la zhi jing bian da ,SNF1he Reg1ke neng tong guo tan dai xie yi zhi yin zi CreAying xiang dian fen mei biao da ,Reg1ji yin qiao chu hou dao zhi he nei CreAhan liang xia jiang ,you dao dian fen shui jie mei de biao da fen bi ,er snf1ji yin qiao chu hou dao zhi he nei CreAhan liang shang sheng ,yi zhi dian fen shui jie mei de biao da fen bi 。guo jiao 、wei jing qian wei su duo tang pei yang ji shang ,Δsnf1he ΔReg1sheng chang su du chu xian cha yi dou cheng xian xia jiang qu shi ,SNF1he Reg1tong guo ji ta tu jing can yu bao wai shui jie mei biao da diao kong 。（3）dui tu bian zhu Δsnf1he ΔReg1huang qu mei du su B1（AFB1）chan liang fen xi fa xian ,AFB1chan liang jun xia jiang ,jiang di lv fen bie wei 80.2%,77.4%（yi chan du pei yang ji YESwei li ）,biao ming SNF1he Reg1dui du su AFB1ge cheng ju you zheng diao kong zuo yong 。（4）fen xi tu bian zhu Δsnf1he ΔReg1du su ge cheng xiang guan ji yin aflD、aflM、aflO、aflPhe zhuai lu yin zi aaflS/flRji yin he diao kong fen sheng bao zi zhuai lu ji yin brlA、abaAde zhuai lu shui ping ke yi zhi dao :snf1、Reg1ji yin qiao chu hou xia diao zhuai lu yin zi aflS/aflRde zhuai lu biao da shui ping ,jin er jie gou ji yin zhuai lu shou dao yi zhi ,zui zhong yin qi du su ge cheng shou zu 。zai aflS/aflRqi dong zi ou cun zai CreAjie ge wei dian ,SNF1he Reg1que shi hou ying xiang CreAlin suan hua shui ping ,rao luan CreAzai he zhi jian de dong tai ping heng ,ying xiang zhuai lu yin zi aflS/aflRde zhuai lu biao da shui ping 。zai Δsnf1he ΔReg1tu bian ti zhong ,can yu diao kong fen sheng bao zi xing cheng fa yo ji yin cheng xian xia diao 。snf1ji yin que shi yin qi fen sheng bao zi zhi hou fa yo xing cheng ,jie ge xian wei guan cha fa xian fen sheng bao zi que xian 。yin ci wo men tui duan ,snf1diao kong fen sheng bao zi de zhun bei jie duan ,gai ji yin que shi dao zhi sheng chang fen hua xiang guan ji yin xia diao ,ru brlA、abaAzhuai lu yi zhi zao cheng fen sheng bao zi zhi hou fa yo 。（5）cai yong qin ran hua sheng shi yan ,yan jiu huang qu mei SNF1he Reg1que shi dui huang qu mei qin ran ding shi li de ying xiang 。yan jiu fa xian ,snf1que shi hou hua sheng biao mian ji hu mei you huang qu mei sheng chang ,huang qu mei qin ran ding shi shou dao ji da cheng du yi zhi ,bing ju jian ce bu dao AFB1。Reg1que shi hou hua sheng shao bu fen biao mian bei huang qu mei bao guo ,sheng chang shou dao yi zhi ,AFB1han liang ji ju xia jiang 。biao ming SNF1he Reg1dui yu huang qu mei qin ran ding shi neng li ju you zheng diao kong zuo yong 。SNF1he Reg1ying xiang shui jie mei ,ru dian fen mei 、guo jiao mei deng de biao da fen bi ,zui zhong xiao ruo huang qu mei qin ran he ding shi neng li ,bi jiao er yan SNF1que shi dui huang qu mei qin ran ding shi neng li zao cheng de ying xiang geng da 。ben yan jiu jie guo wei huang qu mei du su wu ran feng xian ping gu ji fang kong ji shu kai fa dian ding yi ding de li lun ji chu 。
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论文作者分别是来自中国农业科学院的王平，发表于刊物中国农业科学院2019-07-05论文，是一篇关于碳代谢阻遏论文,黄曲霉毒素合成论文,生长发育论文,中国农业科学院2019-07-05论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自中国农业科学院2019-07-05论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。