大麦不同氮利用效率品种筛选及GS2基因的表达分析
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摘要
为探明大麦品种的不同氮利用效率差异及质体型谷氨酰胺合成酶基因的表达差异,以48份大麦品种为材料,并利用不同氮水平营养液水培8周,研究其在低氮(0.4 mmol·L~(-1))、中氮(2.0 mmol·L~(-1))及高氮(5.0 mmol·L~(-1))下根、茎、叶的相关性状,筛选氮高效利用和氮低效利用的大麦品种,并利用qRT-PCR对筛选到的大麦氮高效和氮低效品种的GS2基因表达特性进行分析。结果表明,中氮下,除根干重和根含氮量外,其余的大麦苗期根、茎、叶各生物性状在品种间差异极显著,根据不同氮处理下的48份大麦品种苗期性状以及氮利用效率,筛选到1个氮高效品种(Z0099001)和2个氮低效品种(ARr-91和甘啤7号)。3个氮处理水平下,氮高效品种GS2基因的相对表达量高于氮低效品种,并且氮高效品种和氮低效品种根、茎、叶中GS2基因的相对表达量也各不相同,其中,叶中最高,茎中次之,根中最低。低氮条件下,根茎叶中,Z0099001的GS2基因的相对表达量高于甘啤7号,ARr-91最低。中氮条件下,根、茎、叶中均表现为氮高效品种高于氮低效品种,但在根中差异不显著,茎和叶中差异显著。高氮条件下,3个品种在根中GS2基因的相对表达量较低;在茎和叶中,Z0099001的相对表达量显著高于ARr-91和甘啤7号,且Z0099001叶中GS2基因的相对表达量在所有性状中最高。本研究结果为进一步解析大麦氮高效利用率奠定了基础。
To explore the difference of different nitrogen use efficiency and the glutamine synthase gene( GS2) relative expression in barleycultivars. 48 barley cultivars had been conducted by different nitrogen solution for 8 weeks,the characters related to roots,stems and leaves under low nitrogen( 0. 4 mmol·L~(-1)),normal nitrogen( 2. 0 mmol·L~(-1))and high nitrogen( 5. 0 mmol·L~(-1)) supplies were analyzed,and the high and low nitrogen efficient cultivars were selected. Then relative expression of GS2 in high and low nitrogen efficient cultivars was analyzed by qRT-PCR. The results showed that,1) the differences of seedling roots,stems and leaves among varieties were the most significant in the normal nitrogen condition. Based on seedling characters and nitrogen use efficiency of 48 barley cultivars with different nitrogen treatments,high nitrogen efficient cultivar( Z0099001) and low nitrogen efficient cultivars( ARr-91 and Ganpi7) were obtained. 2) In the three nitrogen treatments,relative expression of GS2 of high nitrogen efficient cultivar was higher than that of the low nitrogen efficient cultivars,and it was also different expressed in roots,stems and leaves,the relative expression was highest in leaves,followed by the stems and the lowest was in roots. For,relative expression of GS2 in roots,stems and leaves of the high nitrogen efficient cultivar Z0099001 was higher than that of low nitrogen efficient cultivar Ganpi7,the low nitrogen efficient cultivar ARr-91 was the lowest in the low nitrogen condition. Under the normal nitrogen condition,it was showed that relative expression of GS2 in roots,stems and leaves of the high nitrogen efficient cultivar was higher than that of the low nitrogen efficient cultivars,but there was no significant difference in roots,the difference was significant in stems and leaves. In the high nitrogen condition,relative expression of GS2 in roots was lower in three cultivars,for stems and leaves,relative expression of GS2 of Z0099001 was obviously higher than that of Ganpi7 and ARr-91,and relative expression of GS2 of Z0099001 in leaves was highest for all characters. The results of this study laid a foundation for the further research of high nitrogen efficient utilization in barley.
To explore the difference of different nitrogen use efficiency and the glutamine synthase gene( GS2) relative expression in barleycultivars. 48 barley cultivars had been conducted by different nitrogen solution for 8 weeks,the characters related to roots,stems and leaves under low nitrogen( 0. 4 mmol·L~(-1)),normal nitrogen( 2. 0 mmol·L~(-1))and high nitrogen( 5. 0 mmol·L~(-1)) supplies were analyzed,and the high and low nitrogen efficient cultivars were selected. Then relative expression of GS2 in high and low nitrogen efficient cultivars was analyzed by qRT-PCR. The results showed that,1) the differences of seedling roots,stems and leaves among varieties were the most significant in the normal nitrogen condition. Based on seedling characters and nitrogen use efficiency of 48 barley cultivars with different nitrogen treatments,high nitrogen efficient cultivar( Z0099001) and low nitrogen efficient cultivars( ARr-91 and Ganpi7) were obtained. 2) In the three nitrogen treatments,relative expression of GS2 of high nitrogen efficient cultivar was higher than that of the low nitrogen efficient cultivars,and it was also different expressed in roots,stems and leaves,the relative expression was highest in leaves,followed by the stems and the lowest was in roots. For,relative expression of GS2 in roots,stems and leaves of the high nitrogen efficient cultivar Z0099001 was higher than that of low nitrogen efficient cultivar Ganpi7,the low nitrogen efficient cultivar ARr-91 was the lowest in the low nitrogen condition. Under the normal nitrogen condition,it was showed that relative expression of GS2 in roots,stems and leaves of the high nitrogen efficient cultivar was higher than that of the low nitrogen efficient cultivars,but there was no significant difference in roots,the difference was significant in stems and leaves. In the high nitrogen condition,relative expression of GS2 in roots was lower in three cultivars,for stems and leaves,relative expression of GS2 of Z0099001 was obviously higher than that of Ganpi7 and ARr-91,and relative expression of GS2 of Z0099001 in leaves was highest for all characters. The results of this study laid a foundation for the further research of high nitrogen efficient utilization in barley.
引文
[1]赖勇,王鹏喜,范贵强,司二静,王晋,杨轲,孟亚雄,李葆春,马小乐,尚勋武,王化俊.大麦SSR标记遗传多样性及其与农艺性状关联分析[J].中国农业科学,2012,46(2):233-242
[2]司二静,张宇,汪军成,孟亚雄,李葆春,马小乐,尚勋武,王化俊.大麦农艺性状与SSR标记的关联分析[J].作物学报,2015,4(7):1064-1072
[3]徐晓明,张迎信,王会民,程式华,曹立勇.水稻氮、磷、钾吸收利用遗传特征研究进展[J].核农学报,2016,30(4):685-694
[4]顾俊荣,董明辉,赵步洪,陈培峰,季红娟,韩立宇.不同水氮管理对水稻干物质积累和茎鞘物质运转及产量的影响[J].核农学报,2016,30(2):347-354
[5]王永华,胡卫丽,李刘霞,段剑钊,郭天财.不同基因型小麦产量和氮利用效率的差异及其相互关系[J].麦类作物学报,2013,33(2):301-308
[6]黄亿,李廷轩,张锡洲,戢林,吴沂珀.氮高效利用基因型大麦氮素转移及氮形态组分特征[J].中国农业科学,2015,48(6):1151-1161
[7]Beato V M,Rexach J,Navarro-Gochicoa M T,Camacho-Cristobal JJ,Herrera-Rodriguez M B,Gonzalez-Fontes A.Boron deficiency increases expressions of asparagine synthetase,glutamate dehydrogenase and glutamine synthetase genes in tobacco roots irrespective of the nitrogen source[J].Soil Science and Plant Nutrition,2014,60(3):314-324
[8]Freeland-Graves J H,Nitzke S.Position of the academy of nutrition and dietetics:total diet approach to healthy eating[J].Journal of the Academy of Nutrition and Dietetics,2013,113(2):307-317
[9]陈云堂,郭东权,杨忠强,张建伟,杨保安,王玉莲.60Coγ射线辐照对水果蔬菜中维生素C的影响[J].核农学报,2009,23(2):302-307
[10]高美须,陈浩,刘春泉,哈益明,陈秀兰.食品辐照技术在中国的研究和商业化应用[J].核农学报,2007,21(6):606-611
[11]Masclaux-Daubresse C,Daniel-Vedele F,Dechorgnat J,Chardon F,Gaufichon L,Suzuki A.Nitrogen uptake,assimilation and remobilization in plants:challenges for sustainable and productive agriculture[J].Annals of Botany,2010,105(7):1141-1157
[12]Xu G,Fan X,Miller A J.Plant nitrogen assimilation and use efficiency[J].Annual Beview of Plant Biology,2012,63(1):153-182
[13]Lin Q,Li C,Zhang C,Peng J,Peng S,Bennett J.Comparative study of immunological properties on glutamine synthetase isozymes in rice plants[J].Acta Botanica Sinica,1999,42(5):471-475
[14]Wang Y,Fu B,Pan L,Chen L,Fu X,Li K.Overexpression of Arabidopsis Dof1,GS1 and GS2 enhanced nitrogen assimilation in transgenic tobacco grown under low-nitrogen conditions[J].Plant Molecular Biology Reporter,2013,31(4):886-900
[15]陆彬彬,周卫,张吉,潘磊,林清华,张楚富.温度对水稻谷氨酰胺合成酶和NADH-谷氨酸合酶表达的影响[J].武汉大学学报,2002,48(2):239-242
[16]王晓波,滕婉,何雪,童依平.大豆谷氨酰胺合成酶基因的分类及根瘤特异表达Gm GS1β2基因功能的初步分析[J].作物学报,2013,39(12):2145-2153
[17]张锡洲,阳显斌,李廷轩,余海英,马程,张世元,罗关友.野生大麦氮素吸收利用的基因型差异[J].核农学报,2011,25(6):1261-1267
[18]陆大雷,刘小兵,赵久然,刘萍,王凤格,陆卫平,张凯迪.甜玉米氮素吸收利用的基因型差异[J].植物营养与肥料学报,2008,14(2):258-263
[19]殷春渊,张庆,魏海燕,张洪程,戴其根,霍中洋,许轲,马群,杭杰,张胜飞.不同产量类型水稻基因型氮素吸收、利用效率的差异[J].中国农业科学,2010,43(1):39-50
[20]黄亿,李廷轩,张锡洲,戢林.氮高效利用基因型大麦的物质生产与氮素积累特性[J].应用生态学报,2014,25(7):1971-1978
[21]魏海燕,张洪程,杭杰,戴其根,霍中洋,许轲,张胜飞,马群,张庆,张军.不同氮素利用效率基因型水稻氮素积累与转移的特性[J].作物学报,2008,34(1):119-125
[22]孙永健,孙园园,徐徽,李玥,严奉君,蒋明金,马均.水氮管理模式对不同氮效率水稻氮素利用特性及产量的影响[J].作物学报,2014,40(9):1639-1649
[23]陈志伟,何婷,陆瑞菊,王亦菲,邹磊,杜志钊,单丽丽,何烨,黄剑华.不同基因型大麦苗期对低氮胁迫的生物学响应[J].上海农业学报,2010,26(1):28-32
[24]Kumada Y,Benson D R,Hillemann D,Hosted T J,Rochefort DA,Thompson C J,Wohlleben W,Tateno Y.Evolution of the glutamine synthetase gene,one of the oldest existing and functioning genes[J].Proceedings of the National Academy of Sciences,1993,90(7):3009-3013
[25]陈志伟,陆瑞菊,王亦菲,何婷,杜志钊,高润红,邹磊,单丽丽,黄剑华.低氮胁迫下大麦谷氨酰胺酶基因的表达分析[J].核农学报,2010,24(6):1182-1184
[26]冯卓,秦智伟,武涛,何红梅.黄瓜细胞质型谷氨酰胺合成酶基因(GS1)的克隆及其在低氮条件下的表达[J].中国农业科学,2012,45(15):3100-3107
[27]初建青,王文艳,房经贵,张春华,张彦平,宋长年.叶面喷施尿素对葡萄氮代谢相关基因表达的影响[J].植物营养与肥料学报,2012,18(2):405-416
[28]Seabra A R,Silva L S,Carvalho H G.Novel aspects of glutamine synthetase(GS)regulation revealed by a detailed expression analysis of the entire GS gene family of Medicago truncatula under different physiological conditions[J].BMC Plant Biology,2013,13(1):1-15
[29]Goodall A J,Kumar P,Tobin A K.Identification and expression analyses of cytosolic glutamine synthetase genes in barley(Hordeum vulgare L.)[J].Plant and Cell Physiology,2013,54(4):492-505
[2]司二静,张宇,汪军成,孟亚雄,李葆春,马小乐,尚勋武,王化俊.大麦农艺性状与SSR标记的关联分析[J].作物学报,2015,4(7):1064-1072
[3]徐晓明,张迎信,王会民,程式华,曹立勇.水稻氮、磷、钾吸收利用遗传特征研究进展[J].核农学报,2016,30(4):685-694
[4]顾俊荣,董明辉,赵步洪,陈培峰,季红娟,韩立宇.不同水氮管理对水稻干物质积累和茎鞘物质运转及产量的影响[J].核农学报,2016,30(2):347-354
[5]王永华,胡卫丽,李刘霞,段剑钊,郭天财.不同基因型小麦产量和氮利用效率的差异及其相互关系[J].麦类作物学报,2013,33(2):301-308
[6]黄亿,李廷轩,张锡洲,戢林,吴沂珀.氮高效利用基因型大麦氮素转移及氮形态组分特征[J].中国农业科学,2015,48(6):1151-1161
[7]Beato V M,Rexach J,Navarro-Gochicoa M T,Camacho-Cristobal JJ,Herrera-Rodriguez M B,Gonzalez-Fontes A.Boron deficiency increases expressions of asparagine synthetase,glutamate dehydrogenase and glutamine synthetase genes in tobacco roots irrespective of the nitrogen source[J].Soil Science and Plant Nutrition,2014,60(3):314-324
[8]Freeland-Graves J H,Nitzke S.Position of the academy of nutrition and dietetics:total diet approach to healthy eating[J].Journal of the Academy of Nutrition and Dietetics,2013,113(2):307-317
[9]陈云堂,郭东权,杨忠强,张建伟,杨保安,王玉莲.60Coγ射线辐照对水果蔬菜中维生素C的影响[J].核农学报,2009,23(2):302-307
[10]高美须,陈浩,刘春泉,哈益明,陈秀兰.食品辐照技术在中国的研究和商业化应用[J].核农学报,2007,21(6):606-611
[11]Masclaux-Daubresse C,Daniel-Vedele F,Dechorgnat J,Chardon F,Gaufichon L,Suzuki A.Nitrogen uptake,assimilation and remobilization in plants:challenges for sustainable and productive agriculture[J].Annals of Botany,2010,105(7):1141-1157
[12]Xu G,Fan X,Miller A J.Plant nitrogen assimilation and use efficiency[J].Annual Beview of Plant Biology,2012,63(1):153-182
[13]Lin Q,Li C,Zhang C,Peng J,Peng S,Bennett J.Comparative study of immunological properties on glutamine synthetase isozymes in rice plants[J].Acta Botanica Sinica,1999,42(5):471-475
[14]Wang Y,Fu B,Pan L,Chen L,Fu X,Li K.Overexpression of Arabidopsis Dof1,GS1 and GS2 enhanced nitrogen assimilation in transgenic tobacco grown under low-nitrogen conditions[J].Plant Molecular Biology Reporter,2013,31(4):886-900
[15]陆彬彬,周卫,张吉,潘磊,林清华,张楚富.温度对水稻谷氨酰胺合成酶和NADH-谷氨酸合酶表达的影响[J].武汉大学学报,2002,48(2):239-242
[16]王晓波,滕婉,何雪,童依平.大豆谷氨酰胺合成酶基因的分类及根瘤特异表达Gm GS1β2基因功能的初步分析[J].作物学报,2013,39(12):2145-2153
[17]张锡洲,阳显斌,李廷轩,余海英,马程,张世元,罗关友.野生大麦氮素吸收利用的基因型差异[J].核农学报,2011,25(6):1261-1267
[18]陆大雷,刘小兵,赵久然,刘萍,王凤格,陆卫平,张凯迪.甜玉米氮素吸收利用的基因型差异[J].植物营养与肥料学报,2008,14(2):258-263
[19]殷春渊,张庆,魏海燕,张洪程,戴其根,霍中洋,许轲,马群,杭杰,张胜飞.不同产量类型水稻基因型氮素吸收、利用效率的差异[J].中国农业科学,2010,43(1):39-50
[20]黄亿,李廷轩,张锡洲,戢林.氮高效利用基因型大麦的物质生产与氮素积累特性[J].应用生态学报,2014,25(7):1971-1978
[21]魏海燕,张洪程,杭杰,戴其根,霍中洋,许轲,张胜飞,马群,张庆,张军.不同氮素利用效率基因型水稻氮素积累与转移的特性[J].作物学报,2008,34(1):119-125
[22]孙永健,孙园园,徐徽,李玥,严奉君,蒋明金,马均.水氮管理模式对不同氮效率水稻氮素利用特性及产量的影响[J].作物学报,2014,40(9):1639-1649
[23]陈志伟,何婷,陆瑞菊,王亦菲,邹磊,杜志钊,单丽丽,何烨,黄剑华.不同基因型大麦苗期对低氮胁迫的生物学响应[J].上海农业学报,2010,26(1):28-32
[24]Kumada Y,Benson D R,Hillemann D,Hosted T J,Rochefort DA,Thompson C J,Wohlleben W,Tateno Y.Evolution of the glutamine synthetase gene,one of the oldest existing and functioning genes[J].Proceedings of the National Academy of Sciences,1993,90(7):3009-3013
[25]陈志伟,陆瑞菊,王亦菲,何婷,杜志钊,高润红,邹磊,单丽丽,黄剑华.低氮胁迫下大麦谷氨酰胺酶基因的表达分析[J].核农学报,2010,24(6):1182-1184
[26]冯卓,秦智伟,武涛,何红梅.黄瓜细胞质型谷氨酰胺合成酶基因(GS1)的克隆及其在低氮条件下的表达[J].中国农业科学,2012,45(15):3100-3107
[27]初建青,王文艳,房经贵,张春华,张彦平,宋长年.叶面喷施尿素对葡萄氮代谢相关基因表达的影响[J].植物营养与肥料学报,2012,18(2):405-416
[28]Seabra A R,Silva L S,Carvalho H G.Novel aspects of glutamine synthetase(GS)regulation revealed by a detailed expression analysis of the entire GS gene family of Medicago truncatula under different physiological conditions[J].BMC Plant Biology,2013,13(1):1-15
[29]Goodall A J,Kumar P,Tobin A K.Identification and expression analyses of cytosolic glutamine synthetase genes in barley(Hordeum vulgare L.)[J].Plant and Cell Physiology,2013,54(4):492-505