甘蓝型油菜G3PDH和PK基因对氮肥的响应
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摘要
为了研究氮肥施用量对油菜含油量以及相关基因表达水平的影响,本研究以甘蓝型油菜‘宁油22号’作为材料,在0 kg/hm~2、320 kg/hm~2、640 kg/hm~2氮肥水平下分别测定‘宁油22号’种子和角果皮的含油量,同时检测G3PDH和PK基因在不同氮肥水平下以及不同组织中的表达含量。结果发现,随着施氮量的增加,‘宁油22号’含油量下降,在0 kg/hm~2和320 kg/hm~2水平时下降不明显,在640 kg/hm~2水平时下降明显,并且随着施氮量的增加,G3PDH和PK基因表达量均增强。本研究还分别分析了在种子和角果皮中G3PDH和PK基因的比值,以及开花30天后跟开花15天后的表达比,结果表明二者均在640 kg/hm~2水平下达到最高;PK基因在种子与角果皮中的比值在640 kg/hm~2水平时达到最高,在开花30天后跟15天后的表达比在0 kg/hm~2水平时达到最高。
In this study,‘Ningyou 22'of cabbage type rapeseed is used as material to study the effect of nitrogen fertilizer application on oil content and gene expression in rapeseed. The oil contents of seed and peel of‘Ningyou 22'were detected at the nitrogen fertilizer levels of 0 kg/hm~2, 320 kg/hm~2 and 640 kg/hm~2,respectively. At the same time, the expressions of G3 PDH and PK genes at different nitrogen fertilizer levels and in different tissues were also detected. The results showed that the oil content of‘Ningyou 22'decreased with the increase of nitrogen application. However, the decrease was not significant at the level of 0 kg/hm~2 and320 kg/hm~2, while it is obvious at the level of 640 kg/hm~2. The expressions of G3 PDH and PK genes were strengthened with the increase of nitrogen application. The ratio ofG3 PDH and PK genes in seed and pericarp was analyzed, as well as the expression ratio after flowering 30 days and 15 days. The results showed that the two ratios reached the highest at the level of 640 kg/hm~2. The ratio of PK genes in seed to pericarp reached the highest at the level of 640 kg/hm~2. And the ratio of PK genes reached the maximum at the level of 0 kg/hm~2 after flowering 30 days and 15 days.
In this study,‘Ningyou 22'of cabbage type rapeseed is used as material to study the effect of nitrogen fertilizer application on oil content and gene expression in rapeseed. The oil contents of seed and peel of‘Ningyou 22'were detected at the nitrogen fertilizer levels of 0 kg/hm~2, 320 kg/hm~2 and 640 kg/hm~2,respectively. At the same time, the expressions of G3 PDH and PK genes at different nitrogen fertilizer levels and in different tissues were also detected. The results showed that the oil content of‘Ningyou 22'decreased with the increase of nitrogen application. However, the decrease was not significant at the level of 0 kg/hm~2 and320 kg/hm~2, while it is obvious at the level of 640 kg/hm~2. The expressions of G3 PDH and PK genes were strengthened with the increase of nitrogen application. The ratio ofG3 PDH and PK genes in seed and pericarp was analyzed, as well as the expression ratio after flowering 30 days and 15 days. The results showed that the two ratios reached the highest at the level of 640 kg/hm~2. The ratio of PK genes in seed to pericarp reached the highest at the level of 640 kg/hm~2. And the ratio of PK genes reached the maximum at the level of 0 kg/hm~2 after flowering 30 days and 15 days.
引文
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[18]闫贵欣,陈碧云,许鲲,等.甘蓝型油菜ACCase、DGAT2和PEP基因对氮素用量的应答[J].植物营养与肥料学报,2012,18(6):1370-1377.
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[20] Stitt M, Mailer C, Matt P, et a1. Steps towards a n integrated view of nitrogen metabolism[J]J.Exp.Bot.2002,53(370):959-970.
[21]张占琴,王金梅,王学军,等.油菜籽粒发育过程中PEPCase活性与油脂、蛋白及亚基积累的特点[J].中国油料作物学报,2009,31(1):14-18.
[22] Hu Y, Wu G, Cao Y, et a1. Breeding response of transcript profiling in developing seeds of Brassica napus[J].BMCMo1.Bio1,2009,1049-1065.
[23] Liu J, Hua W, Yang H L, et al. Effects of specific organs on seed oil accumulation in Brassica napus L.[J].Plant Science,2014,227:60-68.
[2]王汉中.中国油料产业发展的现状,问题和对策[J].中国油料作物学报,2005,27(4):100-105.
[3]钱武,戴成满,郑成彧,等.甘蓝型冬油菜在不同地区生育期,品质和产量差异性分析[J].浙江农业科学,2018,59(5):715-718.
[4]邹小云,陈伦林,李书宇,等.氮、磷、钾、硼肥施用对甘蓝型杂交油菜产量及经济效益的影响[J].中国农业科学, 2011,44(5):917-924.
[5]王荣焕,陈传永,徐田军,等.不同基因型玉米品种产量和氮素利用效率对减施氮肥的响应[J].中国农业科技导报,2018,20(4):101-107.
[6] Kang J M, Turano F J. The putative glutamate receptor 1.1(AtGLR1.1)functions as a regulator of carbon and nitrogen metabolism in Arabidopsis thaliana[J].Proc.Nag.Acta.Sci.USA,2003,100:6872-6877.
[7]周年年.氮素对油菜产量和品质的影响及相关分析研究[D].武汉:华中农业大学,2005.
[8]付三雄,周晓婴,张维,等.种植密度和施氮量对油菜产量、品质及机收性状的影响[J].江苏农业学报,2016,32(3):548-556.
[9]吴金山,张艳,朱传凤,等.4个水稻线粒体丙酮酸激酶基因(mtOsPKs)表达分析[J].热带作物学报,2010,31(5):782-788.
[10]付三雄,张超,陈松,等.甘蓝型油菜丙酮酸激酶基因RNA干扰载体的构建[J].江苏农业学报,2013,29(1):20-27.
[11] Plaxton W C. The functional organization and control of plantrespiration[J].Critical Reviews in Plant Sciences.2006,25:159-198.
[12] Andre C, Froehlich J E, Moll M R, et a1. A heteromericplastidic pyrnvate kinase complex involved in seed oil biosynthesis in Arabidopsis[J]. The Plant Cell,2007,19:2006-2022.
[13]田畅,蒋倩,王枫,等.3个芹菜品种Agnp-G3PDH基因的克隆及其序列和表达特性分析[J].植物资源与环境学报,2014,23(2):1-10.
[14] Chanda B, Xia Y, Mandal M K, et a1. Glycerol-3-phos-phate is a critical mobile inducer of systemic imm unity in plants[J].Nat.Genet,2011,43(5):421-427.
[15] Wang Z X, Zhuge J, Fang H, et a1. Glycerol production by microbial fermentation:a review[J].Biotechno1.Adv.,2001,19(3):201-223.
[16] Vigeolas H, Geigenberger P. Increased levels of glyc-erol-3-phosphate lead to a stimulation of flux into triacylglycerol synthesis after supplying glycerol to developing seeds of Brassica napus L.[J].Planta,2004,219(5):827-835.
[17]张超,付三雄,陈松,等.甘蓝型油菜种子中G3PDH基因特异性表达的ihpRNA载体构建[J].贵州农业科学,2014,42(9):29-34.
[18]闫贵欣,陈碧云,许鲲,等.甘蓝型油菜ACCase、DGAT2和PEP基因对氮素用量的应答[J].植物营养与肥料学报,2012,18(6):1370-1377.
[19]刘后利.实用油菜栽培学[M].上海:上海科学技术出版社,1987.
[20] Stitt M, Mailer C, Matt P, et a1. Steps towards a n integrated view of nitrogen metabolism[J]J.Exp.Bot.2002,53(370):959-970.
[21]张占琴,王金梅,王学军,等.油菜籽粒发育过程中PEPCase活性与油脂、蛋白及亚基积累的特点[J].中国油料作物学报,2009,31(1):14-18.
[22] Hu Y, Wu G, Cao Y, et a1. Breeding response of transcript profiling in developing seeds of Brassica napus[J].BMCMo1.Bio1,2009,1049-1065.
[23] Liu J, Hua W, Yang H L, et al. Effects of specific organs on seed oil accumulation in Brassica napus L.[J].Plant Science,2014,227:60-68.