GLKs转录因子调控番茄果实品质的研究
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摘要
以AC番茄及转化GLKs(Golden-Like)转录因子的AC转基因后代(过表达SlGLK2植株,共抑制SlGLK1植株和共抑制SlGLK2植株)为研究材料,通过对各番茄材料的表型观察、未成熟果实外表皮叶绿素的检测和果实叶绿体发育相关基因SlGLK1、SlGLK2、TKN2(Transcription Factor of Knotted Like Gene)、TKN4和APRR2(Arabidopsis Pseudo Response Regulator 2-Like)的表达分析,以及对成熟果实可溶性固形物、总糖含量进行检测,分析GLKs转录因子对番茄果实品质的影响。结果显示:(1)在表型方面,共抑制SlGLK1基因植株的叶片颜色明显变浅,过表达SlGLK2果实的颜色明显变为深绿色,与之相对应,共抑制SlGLK1基因植株果实顶部叶绿素的含量最低,过表达SlGLK2的果实底部叶绿素含量是AC的5～6倍。(2)过表达SlGLK2转录因子的AC后代成熟果实中可溶性固形物的含量提高了0.3%,总糖含量提高了5.34mg/g。(3)叶绿体发育相关基因的荧光定量PCR显示,与AC相比,转基因材料中SlGLK1和SlGLK2基因的表达量显著提高5倍以上。研究推测,TKN2可能位于SlGLK2基因上游,并调控SlGLK2基因果表达。研究表明,SlGLKs转录因子通过调控番茄叶片和果实叶绿素的合成,加深叶片和未成熟果实的绿色,提高果实内可溶性固形物和总糖的含量,对改善番茄果实的表型和营养品质有重要作用。
Golden-like (GLK)transcription factors play an important role in the growth and formation process of plant leaves and fruit chloroplast.In this study,Solanum lycopersicum(AC)transformed with SlGLK1 and SlGLK2 genes(ACCS SlGLK1,ACCS SlGLK2 and ACOE SlGLK2)was used as the research material,to detect the content of chlorophyll and the expression analysis of related transcription factor genes such as SlGLK1,SlGLK2,TKN2(Transcription Factor of Knotted Like Gene),TKN4(Transcription Factor of Knotted Like Gene)and APRR2(Arabidopsis Pseudo Response Regulator 2-Like)in different developmental stages of green fruits,BRIX and total sugar in mature transgenic fruits.The results showed that:(1)the leaf color in co-suppressed SlGLK1 gene plants became obviously lighter,and the fruit color of overexpressed SlGLK2 turned dark green,which corresponded to the detection of chlorophyll content in fruit.The chlorophyll content in the top of co-suppressed SlGLK1 fruit was the lowest in all materials,but the bottom of fruits were 5-6 times higher in over-expressed SlGLK2 than that of AC.(2)0.3% of BRIX and 5.34 mg/g of total sugar has been increased in over-expressed SlGLK2 gene fruits,respectively.(3)The qPCR of SlGLK1 and SlGLK2 genes showed that the expression in the transgenic materials was significantly increased more than 5 times,even in the co-suppressed transgenic lines.TKN2 might be located upstream of the SlGLK2 gene and regulate SlGLK2 transcription factor.Therefore,SlGLKs transcription factor played an important role in improving the quality of tomato fruit by regulating the synthesis of chlorophyll in leaves and fruits,especially in enhancing the green of tomato leaves and fruit and increasing the BRIX and total sugar in mature fruits.
Golden-like (GLK)transcription factors play an important role in the growth and formation process of plant leaves and fruit chloroplast.In this study,Solanum lycopersicum(AC)transformed with SlGLK1 and SlGLK2 genes(ACCS SlGLK1,ACCS SlGLK2 and ACOE SlGLK2)was used as the research material,to detect the content of chlorophyll and the expression analysis of related transcription factor genes such as SlGLK1,SlGLK2,TKN2(Transcription Factor of Knotted Like Gene),TKN4(Transcription Factor of Knotted Like Gene)and APRR2(Arabidopsis Pseudo Response Regulator 2-Like)in different developmental stages of green fruits,BRIX and total sugar in mature transgenic fruits.The results showed that:(1)the leaf color in co-suppressed SlGLK1 gene plants became obviously lighter,and the fruit color of overexpressed SlGLK2 turned dark green,which corresponded to the detection of chlorophyll content in fruit.The chlorophyll content in the top of co-suppressed SlGLK1 fruit was the lowest in all materials,but the bottom of fruits were 5-6 times higher in over-expressed SlGLK2 than that of AC.(2)0.3% of BRIX and 5.34 mg/g of total sugar has been increased in over-expressed SlGLK2 gene fruits,respectively.(3)The qPCR of SlGLK1 and SlGLK2 genes showed that the expression in the transgenic materials was significantly increased more than 5 times,even in the co-suppressed transgenic lines.TKN2 might be located upstream of the SlGLK2 gene and regulate SlGLK2 transcription factor.Therefore,SlGLKs transcription factor played an important role in improving the quality of tomato fruit by regulating the synthesis of chlorophyll in leaves and fruits,especially in enhancing the green of tomato leaves and fruit and increasing the BRIX and total sugar in mature fruits.
引文
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[9]NAKAMURA H,MURAMATSU M,HAKATA M,et al.Ectopic overexpression of the transcription factor OsGLK1induces chloroplast development in non-green rice cells[J].Plant Cell Physiology,2009,50(11):1 933-1 949.
[10]WATERS M T,WANG P,et al.GLKtranscription factors coordinate expression of the photosynthetic apparatus in Arabidopsis[J].Plant Cell,2009,21(4):1 109-1 128.
[11]POWELL A L T,NGUYEN C V,HILL T,et al.Uniform ripening encodes a Golden2-like transcription factor regulating tomato fruit chloroplast development[J].Science,2012,336(6 089):1 711-1 715.
[12]NADAKUDUTI S S,HOLDSWORTH W L,et al.KNOXgenes influence a gradient of fruit chloroplast development through regulation of GOLDEN2-LIKEexpression in tomato[J].The Plant Journal,2014,78(6):1 022-1 033.
[13]BLANCO-ULATE B,VINCENTI E,et al.Tomato transcriptome and mutant analyses suggest a role for plant stress hormones in the interaction between fruit and Botyytiscinerea[J].Frontiers in Plant Science,2013,4(142):1-16.
[14]WAN S H,ZHOU C M,CHEN Q.Effects of different soil nutrient on the soluble solids content of cherry tomato[J].Agricultural Sciences and Technology,2015,16(1):136-139.
[15]HETHERINGTON S,SMILLIE R,DAVIES W.Photosynthetic activities of vegetative and fruiting tissues of tomato[J].Journal of Experimental Botany,1998,49(324):1 173-1 181.
[2]汪莹,孔俊花,陈微微,等.番茄果实成熟相关转录因子的研究进展[J].园艺学报,2014,41(9):1 811-1 820.WANGY,KONG J H,CHEN W W,et al.Fruit ripening-related transcription factors in tomato[J].Acta Horticulturae Sinica,2014,41(9):1 811-1 820.
[3]范中奇,邝健飞,陆旺金,等.转录因子调控果实成熟和衰老机制研究进展[J].园艺学报,2015,42(9):1 649-1 663.FAN Z Q,KUANG J F,LU W J,et al.Advances in research of the mechanism of transcription factors involving in regulating fruit ripening and senescence[J].Acta Horticulturae Sinica,2015,42(9):1 649-1 663.
[4]刘俊芳,张佳,李贺,等.植物GOLGEN2-Like转录因子研究进展[J].分子植物育种,2017,15(10):3 949-3 956.LIU J F,ZHANG J,LI H,et al.Research progress of plant GOLDEN2-Like transcription factor[J].Molecular Plant Breeding,2017,15(10):3 949-3 956.
[5]ROSSINI L,CRIBB L,MARTIN D J,et al.The maize golden2gene defines a novel class of transcriptional regulators in plants[J].Plant Cell,2001,13(5):1 231-1 244.
[6]FITTER D W,MARTIN D J,COPLEY M J,et al.GLKgene pairs regulate chloroplast development in diverse plant species[J].The Plant Journal,2002,31(6):713-727.
[7]BTAND A,BOROVSKY Y,et al.CaGLK2regulates natural variation of chlorophyll content and fruit color in pepper fruit[J].Theoretical and Applied Genetics,2014,127(10):2 139-2 148.
[8]YASUMURA Y,MOYLAN E C,LANGDALE J A.A conserved transcription factor mediates nuclear control of organelle biogenesis in anciently diverged land plants[J].Plant Cell,2005,17(7):1 894-1 907.
[9]NAKAMURA H,MURAMATSU M,HAKATA M,et al.Ectopic overexpression of the transcription factor OsGLK1induces chloroplast development in non-green rice cells[J].Plant Cell Physiology,2009,50(11):1 933-1 949.
[10]WATERS M T,WANG P,et al.GLKtranscription factors coordinate expression of the photosynthetic apparatus in Arabidopsis[J].Plant Cell,2009,21(4):1 109-1 128.
[11]POWELL A L T,NGUYEN C V,HILL T,et al.Uniform ripening encodes a Golden2-like transcription factor regulating tomato fruit chloroplast development[J].Science,2012,336(6 089):1 711-1 715.
[12]NADAKUDUTI S S,HOLDSWORTH W L,et al.KNOXgenes influence a gradient of fruit chloroplast development through regulation of GOLDEN2-LIKEexpression in tomato[J].The Plant Journal,2014,78(6):1 022-1 033.
[13]BLANCO-ULATE B,VINCENTI E,et al.Tomato transcriptome and mutant analyses suggest a role for plant stress hormones in the interaction between fruit and Botyytiscinerea[J].Frontiers in Plant Science,2013,4(142):1-16.
[14]WAN S H,ZHOU C M,CHEN Q.Effects of different soil nutrient on the soluble solids content of cherry tomato[J].Agricultural Sciences and Technology,2015,16(1):136-139.
[15]HETHERINGTON S,SMILLIE R,DAVIES W.Photosynthetic activities of vegetative and fruiting tissues of tomato[J].Journal of Experimental Botany,1998,49(324):1 173-1 181.