‘黄金梨’缺铁黄化叶片受GA3诱导复绿的机理研究

doi:10.16420/j.issn.0513-353x.2020-0285

摘要/Abstract

摘要：

以‘黄金梨’缺铁黄化植株与正常植株为试材,于生长期黄化植株叶面喷施50、100、200和400 mg · L^-1GA₃溶液,测定叶片中Fe、GA₃、GA₄、IAA和JA含量,对叶绿体进行了电镜显微结构观察,并运用荧光定量PCR技术分析了Fe代谢与GA信号转导相关基因的表达量。结果表明,外源 GA₃处理后9 d,黄化叶均有不同程度复绿,叶脉复绿明显,叶身部分复绿,其中400 mg · L^-1 GA₃处理后12 d黄化叶复绿最明显。处理后9和12 d缺铁黄化叶片Fe含量显著高于清水处理对照。外源GA₃能诱导黄化叶片内FER1、FER2、FER3、FRO2、IRT1和FD1的表达,其中12 d各处理FER2和IRT1基因的表达量分别是对照10倍和40倍以上,说明外源GA₃促进了叶片内铁的贮藏、还原与转运相关基因的表达。外源GA₃处理显著提高了叶片中内源GA₃和GA₄含量,迅速诱导GID11和GID13基因的表达,其中50 mg · L^-1 GA₃处理,GID11表达量在3 d是对照的76.45倍。400 mg · L^-1 GA₃处理后12 d的叶片类囊体基粒片层较黄化叶片清晰,断裂片层少。试验结果初步揭示了GA₃诱导‘黄金梨’黄化叶复绿的机理。

关键词: 梨, 叶片复绿, Fe代谢, GA信号转导基因, qRT-PCR

Abstract:

In order to reveal the regreening effect and physiological mechanism of the iron-deficiency chlorosis leaves induced by GA₃,the iron-deficiency chlorosis and normal plants of‘Whangkeumbae’（Pyrus pyrifolia Nakai）were used as the experiment materials,and GA₃ solutions of 50,100,200 and 400 mg · L^-1 was sprayed on chlorosis leaves during the growth period. The contents of Fe,GA₃,GA₄,IAA and JA in leaves were determined,the chloroplast ultrastructure was observed and photographed under an electron microscope,and the expression levels of genes related to Fe metabolism or GA signal transduction were analyzed by fluorescence quantitative PCR. The results showed that on the 9th day after treatment with exogenous GA₃,the chlorosis leaves showed different degrees of regreening,with obvious vein regreening and partial body regreening. On the 12th day of treatment with 400 mg · L^-1 GA₃,the regreening of chlorosis leaves were the most obvious. The Fe content in regreening leaves were significantly higher than that in control with water on the 9th and 12th day after GA₃treatment. Exogenous GA₃ could induce the expressions of FER1,FER2,FER3,FRO2,IRT1 and FD1 in chlorosis leaves. On the 12th day,the expression levels of FER2 and IRT1 in each treatment were more than 10 and 40 times higher than that in the control group,indicating that exogenous GA₃ promoted the expression of genes related to iron storage,reduction and transport in leaves. Exogenous GA₃ treatment significantly increased the contents of endogenous GA₃ and GA₄in leaves and rapidly induced the expression of GID11 and GID13. Among them,the expression level of GID11 was 76.45 times higher than that of the control group on the 3rd day after the treatment with 50 mg · L^-1 GA₃. On the 12th day after treatment with 400 mg · L^-1 GA₃,the thylakoid granule lamellae in leaves were clearer than that of chlorosis leaves,with fewer fractured lamellae. The results preliminarily revealed regreening mechanism of chlorosis leaves induced by GA₃in‘Whangkeumbae’.

Key words: pear, leaf regreening, Fe metabolism, GA transduction gene, qRT-PCR

中图分类号:

S661.2

贾兵, 郭国凌, 王友煜, 叶振风, 刘莉, 刘普, 衡伟, 朱立武. ‘黄金梨’缺铁黄化叶片受GA₃诱导复绿的机理研究[J]. 园艺学报, 2021, 48(2): 254-264.

JIA Bing, GUO Guoling, WANG Youyu, YE Zhenfeng, LIU Li, LIU Pu, HENG Wei, ZHU Liwu. Studies on the Regreening Mechanism of the Iron-deficiency Chlorosis Leaves induced by GA₃ in‘Whangkeumbae’（Pyrus pyrifolia Nakai）[J]. Acta Horticulturae Sinica, 2021, 48(2): 254-264.

图/表 7

表1 实时荧光定量PCR引物序列

Table 1 The primer sequences for RT-qPCR

基因 Gene	引物序列 Primer sequence	基因 Gene	引物序列 Primer sequence
FER1	Forword：AGGTCTCTCTTGCCCGTCAG	GA2ox	Forword：AATCGCTCTGTTCATCC
FER1	Reverse：GCAACGTTGTCCCTGTCAAA		Reverse：GGAAATAACGCAGAAGAG
FER2	Forword：TTCTCGCCTCTGACCTCCTC	DELLA4	Forword：ACAGCATACAGACCCGTCCAT
FER2	Reverse：CGAAATAGGCGTACATGGCA	DELLA4	Reverse：TGTACCCCAACATCGCCAGAA
FER3	Forword：TGCTTTTGTTGTCCCTGTGG	GID11	Forword：AGCGGAATTCCTTGACCGGAA
FER3	Reverse：TCCTCTTTCTTCCTCGCTCG	GID11	Reverse：CACCGCTCTCTCAGGATCAACA
FRO2	Forword：TCATTGCGGTGTTGGGTTGT	GID13	Forword：TTCCGGTTGACGGGGTTTTCT
FRO2	Reverse：ATCGCTTCTTCTTTCTGTCATTGCT	GID13	Reverse：GAAGCTTCCGCCGTGGAAAAA
IRT1	Forword：GTCTGAGGAGGACGAGTGC	GID15	Forword：ATCATCGGGGGACGATTCGAT
IRT1	Reverse：TGGAAGGCTGGTATGGAAC	GID15	Reverse：TCATCCTGGTCAACGCAGTCA
FD1	Forword：GGTCAGCACCTCCTTCATCC	GID16	Forword：TCGAGCCAGAACGTGGTTGAA
FD1	Reverse：TACGTCCTCAGGGCACTCAA	GID16	Reverse：TGCATTCAGAACCATGGCAGC

图1 ‘黄金梨’黄化植株喷施GA3后叶片复绿情况

Fig. 1 Regreening performance of the chlorosis leaves treated by GA3 solutions on‘Hwangkumbae’pear

图2 外源GA3处理对叶片中Fe含量的影响

Fig. 2 The Fe content in leaves with the exogenous GA3 solutions treatment

图3 外源GA3处理对铁代谢相关基因表达的影响

Fig. 3 Effects of the exogenous GA3 treatment on the expression of the genes related to iron metabolism

图4 外源GA3处理对叶片内源激素含量的影响

Fig. 4 Effects of the exogenous GA3 treatment on the expression of the genes related to iron metabolism

图5 外源GA3处理对其信号传导相关基因表达的影响

Fig. 5 Effects of the exogenous GA3 treatment on the related genes expression for GA signal transduction

图6 外源GA3处理对叶绿体超微结构的影响 A. 正常叶片中叶绿体片层结构电镜图;B. 黄化叶片中叶绿体片层结构电镜图;C. 400 mg · L-1 GA3处理后12 d叶片叶绿体片层结构电镜图。

Fig. 6 Effect of exogenous GA3 treatment on chloroplast ultrastructure A. The grana lamellae ultrastructure of chloroplast in normal leaves;B. The grana lamellae ultrastructure of chloroplast in chlorosis leaves;C. The grana lamellae ultrastructure of chloroplast in chlorosis leaves after treated with 400 mg · L-1 GA3 12 days.

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‘黄金梨’缺铁黄化叶片受GA₃诱导复绿的机理研究

Studies on the Regreening Mechanism of the Iron-deficiency Chlorosis Leaves induced by GA₃ in‘Whangkeumbae’（Pyrus pyrifolia Nakai）

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