本文主要研究内容
作者康亮（2019）在《不同氮效率木薯根系生长、光合作用及转录组分析》一文中研究指出：木薯(Manihot esculenta Crantz)系大戟科(Euphorbiaceae)木薯属(Manihot)多年生灌木植物,主要栽培在热带和亚热带地区,是三大薯类之一。在我国,木薯不仅是主要的淀粉工业和生物质能源原料,也是粮食安全的有效补充作物。氮素是木薯正常生长发育过程的首要元素,一定程度的增施氮肥能提高木薯产量,但过量施氮既浪费资源又带来一系列的环境污染问题。因此,利用遗传改良手段,选育氮高效作物品种被认为是提高氮肥利用效率、保护生态环境和促进农业可持续发展的有效途径。本研究首先以25个木薯品种为材料,通过2年的大田试验并以产量和相对氮积累量对其氮效率进行分类。在此基础上,从氮高效型和氮低效型品种中分别选出代表品种华南10号(SC10)和华南205号(SC205),在低氮、常氮两种供氮水平下,通过分析比较两个品种间的氮积累特征、根系形态、根系构型和硝态氮吸收动力学差异,以及两个品种间的干物质积累特征、光合特性和氮代谢同化酶差异等,从氮素吸收和利用两个方面解释木薯氮高效的机理。最后利用RNA-Seq技术对两因素(品种X处理)下木薯功能叶的基因表达进行探测,通过生物信息学对这些差异基因的生物学功能和代谢途径进行深入研究,了解氮高效基因型在氮素吸收利用过程中可能的分子途径。主要研究结果如下:1.木薯氮效率的评价供试25个木薯品种的氮素吸收(吸氮量)和氮素利用效率在品种、氮处理及交互作用上存在极显著差异,可从中筛选出氮高效品种。以低氮处理产量和相对氮积累量(低氮/高氮)为指标,将25个木薯品种分为四个类型,通过比较两年的分类情况,最终选择SC10作为氮高效型品种,SC205作为氮低效型品种。通径分析表明,木薯的吸收效率和氮素利用效率均对整株干物重有正的贡献作用,且吸收效率的贡献率较高。2.不同氮效率木薯品种氮素吸收差异特征(1)不同氮效率木薯品种各生育期氮素积累量差异显著;不同阶段的氮积累量结果表明,SC10在苗期至块根形成期(即出苗后0～80天)氮素积累量显著高于SC205,N1、NO条件下分别达1.42和1.56倍;而在块根形成期至膨大期(出苗后80～160天)两个木薯品种的氮素积累量无显著差异。表明,两种氮效率型木薯品种的氮素吸收差异在块根形成期之前存在,之后差异变小。(2)苗期SC10的总根长、根系表面积和根系体积显著高于SC205;两品种的细根(直径<0.5mm)比例在N1条件下无显著差异,但NO条件下SC10在细根、中根(直径0.5～1.5mm)的分配比例显著高于SC205。相关分析表明,木薯的整株吸氮量与总根长及单位根长吸氮速率呈显著正相关,进一步分析表明,单位根长吸氮速率对于整株吸氮量的贡献大于总根长。(3)微根管试验发现,除出苗30天后,其余各时期SC10在浅层(35cm)、深层(65cm)土壤的根长密度大于SC205,但差异不显著。玻璃窗观察法同样证实了SC10扎根更深,深层根分布比例更高,其中出苗180天后,SC10在80～180cm处的分布比例在N1、NO条件下比SC205高10.599%、3.5%;SC205的根系则较集中在浅层土壤中。(4)根系N03-吸收动力学试验表明SC10氮高效木薯拥有较低的Km值及Vm值,而SC205氮低效木薯的Km值及Vm值均较高。表明氮高效型木薯品种更适应低氮条件的外界环境。3.不同氮效率木薯品种氮素利用差异特征(1)两种供氮水平下SC10的全生育期干物质量均显著高于SC205。干物质合成强度结果表明,块根形成期至膨大期的干物质合成强度最高,且SC10在各个生育阶段的干物质合成强度在N1条件下分别比SC205高25.4%、35.6%、36.5%、31.0%。表明氮高效型木薯品种的干物质合成能力强于氮低效型木薯品种。(2)在木薯的各生育期,SC10叶面积指数显著高于SC205,但两个木薯品种间的净光合速率(Pn)、气孔导度(Gs)、胞间二氧化碳(Ci)、蒸腾速率(Tm)则随氮处理和生育期进程互有高低。最大光量子产量(Fv/Fm)在块根形成期和膨大期呈现SC205显著高于SC10,实际光量子产量(Y(Ⅱ))值则在块根形成期呈现SC205显著高于SC10。相关分析表明,在全生育期内叶面积指数始终与干物质量极显著正相关,而Pn值仅在块根形成期与干物质量显著正相关,通径分析表明,叶面积指数对干物质量积累的贡献作用远大于Pn值。(3)两种供氮水平下木薯叶片的NR、GS、GOGAT、GDH活性存在显著的品种间差异,SC10显著高于SC205,说明氮高效型木薯品种对的N03-同化速率较高。通过相关分析及通径分析上述4种酶活性与木薯吸氮量的关系发现,对于氮高效型品种SC10,NR活性对其吸氮量的贡献率最高,而对于氮低效型品种SC205,GOGAT活性对其吸氮量的贡献最高,表明不同氮效率型木薯品种存在不同的氮同化酶限制。4.不同氮效率型木薯品种叶片的差异表达基因利用RNA-Seq技术获得了两种供氮水平下两个品种叶片的转录组信息,经组装拼接后,SC10N1、SC10N0、SC205N1、SC205N0分别共有27096、26945、27598、24928个基因表达。利用DGE方法分析了SC10、SC205在N0处理下的差异表达,分别得到68、1307个差异表达,说明N0处理对SC10的影响较小。GO分析显示,在N0处理下,SC205上调基因显著富集于氧化还原过程中,下调基因富集于脂类代谢、光合系统中,表明SC205光合体系受损严重。KEGG富集分析显示,在N0处理下,SC10上调基因显著富集于类黄酮代谢途径,该途径与色素及抗氧化活性有关,而SC205下调基因显著富集于光合作用和天线蛋白中,进一步说明SC205的光合体系遭到破坏。
Abstract
mu shu (Manihot esculenta Crantz)ji da ji ke (Euphorbiaceae)mu shu shu (Manihot)duo nian sheng guan mu zhi wu ,zhu yao zai pei zai re dai he ya re dai de ou ,shi san da shu lei zhi yi 。zai wo guo ,mu shu bu jin shi zhu yao de dian fen gong ye he sheng wu zhi neng yuan yuan liao ,ye shi liang shi an quan de you xiao bu chong zuo wu 。dan su shi mu shu zheng chang sheng chang fa yo guo cheng de shou yao yuan su ,yi ding cheng du de zeng shi dan fei neng di gao mu shu chan liang ,dan guo liang shi dan ji lang fei zi yuan you dai lai yi ji lie de huan jing wu ran wen ti 。yin ci ,li yong wei chuan gai liang shou duan ,shua yo dan gao xiao zuo wu pin chong bei ren wei shi di gao dan fei li yong xiao lv 、bao hu sheng tai huan jing he cu jin nong ye ke chi xu fa zhan de you xiao tu jing 。ben yan jiu shou xian yi 25ge mu shu pin chong wei cai liao ,tong guo 2nian de da tian shi yan bing yi chan liang he xiang dui dan ji lei liang dui ji dan xiao lv jin hang fen lei 。zai ci ji chu shang ,cong dan gao xiao xing he dan di xiao xing pin chong zhong fen bie shua chu dai biao pin chong hua na 10hao (SC10)he hua na 205hao (SC205),zai di dan 、chang dan liang chong gong dan shui ping xia ,tong guo fen xi bi jiao liang ge pin chong jian de dan ji lei te zheng 、gen ji xing tai 、gen ji gou xing he xiao tai dan xi shou dong li xue cha yi ,yi ji liang ge pin chong jian de gan wu zhi ji lei te zheng 、guang ge te xing he dan dai xie tong hua mei cha yi deng ,cong dan su xi shou he li yong liang ge fang mian jie shi mu shu dan gao xiao de ji li 。zui hou li yong RNA-Seqji shu dui liang yin su (pin chong Xchu li )xia mu shu gong neng xie de ji yin biao da jin hang tan ce ,tong guo sheng wu xin xi xue dui zhe xie cha yi ji yin de sheng wu xue gong neng he dai xie tu jing jin hang shen ru yan jiu ,le jie dan gao xiao ji yin xing zai dan su xi shou li yong guo cheng zhong ke neng de fen zi tu jing 。zhu yao yan jiu jie guo ru xia :1.mu shu dan xiao lv de ping jia gong shi 25ge mu shu pin chong de dan su xi shou (xi dan liang )he dan su li yong xiao lv zai pin chong 、dan chu li ji jiao hu zuo yong shang cun zai ji xian zhe cha yi ,ke cong zhong shai shua chu dan gao xiao pin chong 。yi di dan chu li chan liang he xiang dui dan ji lei liang (di dan /gao dan )wei zhi biao ,jiang 25ge mu shu pin chong fen wei si ge lei xing ,tong guo bi jiao liang nian de fen lei qing kuang ,zui zhong shua ze SC10zuo wei dan gao xiao xing pin chong ,SC205zuo wei dan di xiao xing pin chong 。tong jing fen xi biao ming ,mu shu de xi shou xiao lv he dan su li yong xiao lv jun dui zheng zhu gan wu chong you zheng de gong suo zuo yong ,ju xi shou xiao lv de gong suo lv jiao gao 。2.bu tong dan xiao lv mu shu pin chong dan su xi shou cha yi te zheng (1)bu tong dan xiao lv mu shu pin chong ge sheng yo ji dan su ji lei liang cha yi xian zhe ;bu tong jie duan de dan ji lei liang jie guo biao ming ,SC10zai miao ji zhi kuai gen xing cheng ji (ji chu miao hou 0～80tian )dan su ji lei liang xian zhe gao yu SC205,N1、NOtiao jian xia fen bie da 1.42he 1.56bei ;er zai kuai gen xing cheng ji zhi peng da ji (chu miao hou 80～160tian )liang ge mu shu pin chong de dan su ji lei liang mo xian zhe cha yi 。biao ming ,liang chong dan xiao lv xing mu shu pin chong de dan su xi shou cha yi zai kuai gen xing cheng ji zhi qian cun zai ,zhi hou cha yi bian xiao 。(2)miao ji SC10de zong gen chang 、gen ji biao mian ji he gen ji ti ji xian zhe gao yu SC205;liang pin chong de xi gen (zhi jing <0.5mm)bi li zai N1tiao jian xia mo xian zhe cha yi ,dan NOtiao jian xia SC10zai xi gen 、zhong gen (zhi jing 0.5～1.5mm)de fen pei bi li xian zhe gao yu SC205。xiang guan fen xi biao ming ,mu shu de zheng zhu xi dan liang yu zong gen chang ji chan wei gen chang xi dan su lv cheng xian zhe zheng xiang guan ,jin yi bu fen xi biao ming ,chan wei gen chang xi dan su lv dui yu zheng zhu xi dan liang de gong suo da yu zong gen chang 。(3)wei gen guan shi yan fa xian ,chu chu miao 30tian hou ,ji yu ge shi ji SC10zai jian ceng (35cm)、shen ceng (65cm)tu rang de gen chang mi du da yu SC205,dan cha yi bu xian zhe 。bo li chuang guan cha fa tong yang zheng shi le SC10za gen geng shen ,shen ceng gen fen bu bi li geng gao ,ji zhong chu miao 180tian hou ,SC10zai 80～180cmchu de fen bu bi li zai N1、NOtiao jian xia bi SC205gao 10.599%、3.5%;SC205de gen ji ze jiao ji zhong zai jian ceng tu rang zhong 。(4)gen ji N03-xi shou dong li xue shi yan biao ming SC10dan gao xiao mu shu yong you jiao di de Kmzhi ji Vmzhi ,er SC205dan di xiao mu shu de Kmzhi ji Vmzhi jun jiao gao 。biao ming dan gao xiao xing mu shu pin chong geng kuo ying di dan tiao jian de wai jie huan jing 。3.bu tong dan xiao lv mu shu pin chong dan su li yong cha yi te zheng (1)liang chong gong dan shui ping xia SC10de quan sheng yo ji gan wu zhi liang jun xian zhe gao yu SC205。gan wu zhi ge cheng jiang du jie guo biao ming ,kuai gen xing cheng ji zhi peng da ji de gan wu zhi ge cheng jiang du zui gao ,ju SC10zai ge ge sheng yo jie duan de gan wu zhi ge cheng jiang du zai N1tiao jian xia fen bie bi SC205gao 25.4%、35.6%、36.5%、31.0%。biao ming dan gao xiao xing mu shu pin chong de gan wu zhi ge cheng neng li jiang yu dan di xiao xing mu shu pin chong 。(2)zai mu shu de ge sheng yo ji ,SC10xie mian ji zhi shu xian zhe gao yu SC205,dan liang ge mu shu pin chong jian de jing guang ge su lv (Pn)、qi kong dao du (Gs)、bao jian er yang hua tan (Ci)、zheng teng su lv (Tm)ze sui dan chu li he sheng yo ji jin cheng hu you gao di 。zui da guang liang zi chan liang (Fv/Fm)zai kuai gen xing cheng ji he peng da ji cheng xian SC205xian zhe gao yu SC10,shi ji guang liang zi chan liang (Y(Ⅱ))zhi ze zai kuai gen xing cheng ji cheng xian SC205xian zhe gao yu SC10。xiang guan fen xi biao ming ,zai quan sheng yo ji nei xie mian ji zhi shu shi zhong yu gan wu zhi liang ji xian zhe zheng xiang guan ,er Pnzhi jin zai kuai gen xing cheng ji yu gan wu zhi liang xian zhe zheng xiang guan ,tong jing fen xi biao ming ,xie mian ji zhi shu dui gan wu zhi liang ji lei de gong suo zuo yong yuan da yu Pnzhi 。(3)liang chong gong dan shui ping xia mu shu xie pian de NR、GS、GOGAT、GDHhuo xing cun zai xian zhe de pin chong jian cha yi ,SC10xian zhe gao yu SC205,shui ming dan gao xiao xing mu shu pin chong dui de N03-tong hua su lv jiao gao 。tong guo xiang guan fen xi ji tong jing fen xi shang shu 4chong mei huo xing yu mu shu xi dan liang de guan ji fa xian ,dui yu dan gao xiao xing pin chong SC10,NRhuo xing dui ji xi dan liang de gong suo lv zui gao ,er dui yu dan di xiao xing pin chong SC205,GOGAThuo xing dui ji xi dan liang de gong suo zui gao ,biao ming bu tong dan xiao lv xing mu shu pin chong cun zai bu tong de dan tong hua mei xian zhi 。4.bu tong dan xiao lv xing mu shu pin chong xie pian de cha yi biao da ji yin li yong RNA-Seqji shu huo de le liang chong gong dan shui ping xia liang ge pin chong xie pian de zhuai lu zu xin xi ,jing zu zhuang pin jie hou ,SC10N1、SC10N0、SC205N1、SC205N0fen bie gong you 27096、26945、27598、24928ge ji yin biao da 。li yong DGEfang fa fen xi le SC10、SC205zai N0chu li xia de cha yi biao da ,fen bie de dao 68、1307ge cha yi biao da ,shui ming N0chu li dui SC10de ying xiang jiao xiao 。GOfen xi xian shi ,zai N0chu li xia ,SC205shang diao ji yin xian zhe fu ji yu yang hua hai yuan guo cheng zhong ,xia diao ji yin fu ji yu zhi lei dai xie 、guang ge ji tong zhong ,biao ming SC205guang ge ti ji shou sun yan chong 。KEGGfu ji fen xi xian shi ,zai N0chu li xia ,SC10shang diao ji yin xian zhe fu ji yu lei huang tong dai xie tu jing ,gai tu jing yu se su ji kang yang hua huo xing you guan ,er SC205xia diao ji yin xian zhe fu ji yu guang ge zuo yong he tian xian dan bai zhong ,jin yi bu shui ming SC205de guang ge ti ji zao dao po huai 。
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标签：木薯论文; 氮高效论文; 氮素吸收论文; 差异表达论文; 广西大学2019-10-14论文;