2,4-D调控番茄果形的转录基础

doi:10.16420/j.issn.0513-353x.2022-0023

摘要/Abstract

摘要：

番茄冬春季设施生产中为提高坐果率而采用的2,4-D蘸花措施会导致畸形果率增加，其转录层面的机理尚不明确。本研究中对以醋栗番茄LA1589（Solanum pimpinellifolium accession LA1589）为背景的果形近等基因系WT、sun和ovate花蕾喷施2,4-D，而后对花蕾转录组与成熟果实果形进行分析。结果发现，3个近等基因系在2,4-D处理3周后所形成的子房及其发育而来的成熟果实的果梗端均伸长，并最终导致子房与果实的形状指数增大，而在此过程中两个果形位点与2,4-D间存在互作。转录组分析发现，2,4-D处理可能通过激活生长素信号通路，促进生长素极性运输与细胞分裂素合成以及增加ATHB和bHLH转录因子的积累来改变果形；sun与ovate位点通过与2,4-D互作进一步增强了生长素积累、信号转导与极性运输，扩大了受影响的转录因子范围，最终强化了2,4-D对果形的影响；同时该互作还在sun与ovate近等基因系中分别上调了BRX与CKX5基因的表达来进一步增强上述过程；最后，ovate与2,4-D的互作导致两个SWEET基因的表达下调并可能影响糖类向子房中的运输。

关键词: 番茄, 2,4-D, 生长素, 花蕾, 果形, 转录组, sun, ovate

Abstract:

The application of the synthetic auxin，2,4-D，to the inflorescences is often used in winter and spring to promote fruit set of tomato in the greenhouse. However，the improper use of this method leads to the fruit deformity sometimes，of which the mechanism has not been well illustrated yet. Based on the previous research，2,4-D was sprayed to the plants of the WT，sun and ovate NILs in the LA1589 background in this study. Ovaries and fruits from flowers which were anthesis at three weeks after the treatment were investigated at the morphological and histological levels，and meanwhile flower buds were collected for RNA-seq analysis. Results indicated that the elongation of ovaries and fruits induced by the 2,4-D treatment could be attributed to the elongation of the proximal end part. In the above process，interactions were discovered between the investigated fruit shape loci and the 2,4-D treatment. RNA-seq analysis revealed that the effect of 2,4-D on the fruit and ovary shape may through its activation on the auxin signaling，polar transportation as well as its promotion on the cytokinin biosynthesis and the accumulation of the ATHB and bHLH transcription factors. Meanwhile，interactions were also observed between the investigated fruit shape loci and the 2,4-D treatment at the transcriptional level. Those interactions strengthened the accumulation of auxin biosynthesis，signal transduction and polar transportation and impacted the expression of more transcription factor genes. In addition，those interactions also up-regulated the expression of BRX and CKX5 in sun and ovate NILs，respectively，which may further enhance the effects of 2,4-D on ovary and fruit shape. Besides what is mentioned above，in ovate NIL 2,4-D treatment decreased the expression of two SWEET genes which may affect the transportation of sugars into the ovaries.

Key words: tomato, 2,4-D, auxin, flower bud, fruit shape, transcriptome, sun, ovate

中图分类号:

S641.2

徐悦, 李子雄, 陈婕, 孙亮. 2,4-D调控番茄果形的转录基础[J]. 园艺学报, 2023, 50(4): 802-814.

XU Yue, LI Zixiong, CHEN Jie, SUN Liang. Transcriptional Bases of the Regulation of 2,4-D on Tomato Fruit Shape[J]. Acta Horticulturae Sinica, 2023, 50(4): 802-814.

图/表 6

图1 2,4-D处理后果形及野生型近等基因系开花期子房形状变化

Fig. 1 Fruit shape and WT anthesis ovary shape changes after 2,4-D treatment

表1 2,4-D处理后果实表型分析

Table 1 Morphological analysis of fruits after 2,4-D treatment

		2,4-D处理/(mmol · L^-1） 2,4-D treatment	最大宽度/cm Maximum width	最大长度/cm Maximum height	果形指数 Fruit shape index	果梗端比例 Proximal end ratio
基因型 Genotype	WT	0	1.20 ± 0.01 a	1.23 ± 0.01 d	1.02 ± 0.01 d	0.13 ± 0.01 d
	WT	270	0.95 ± 0.03 b	1.35 ± 0.02 cd	1.43 ± 0.03 c	0.31 ± 0.03 bc
	sun	0	1.14 ± 0.04 a	1.65 ± 0.03 abc	1.45 ± 0.03 c	0.15 ± 0.03 d
	sun	270	1.65 ± 0.03 b	1.92 ± 0.03 a	2.48 ± 0.27 a	0.36 ± 0.03 b
	ovate	0	1.19 ± 0.03 a	1.52 ± 0.12 bcd	1.28 ± 0.13 cd	0.21 ± 0.03 cd
	ovate	270	0.92 ± 0.12 b	1.80 ± 0.25 ab	1.96 ± 0.05 b	0.49 ± 0.08 a
因素效应 Factorial effects （P-value）		基因型Genotype	0.02	1.93e-05	1.03e-06	2.44e-4
		处理Treatment	4.56e-07	1.28e-03	4.24e-08	5.36e-08
		基因型与处理互作 Genotype × Treatment	0.31	0.45	2.94e-03	0.17

图2 2,4-D处理后各近等基因系花蕾RNA-seq质控（A）和表达基因数（B）

Fig. 2 RNA-seq sequencing quality of the anthesis ovaries after 2,4-D treatment（A）and number of the expressed genes（B）

图3 2,4-D处理后WT（A）、sun（B）和ovate（C）近等基因系花蕾中差异表达基因热图左侧分数中分子与分母分别表示热图中的上调与下调基因数。

Fig. 3 Heatmaps of the differentially expressed genes（DEGs）in the anthesis ovaries of WT（A），sun（B）and ovate（C）NILs after 2,4-D treatment Numerator and denominator in the left represent the number of the up-regulated and down-regulated genes，respectively.

图4 2,4-D处理后各近等基因系花蕾差异表达基因韦恩图分析及感兴趣基因表达热图 A：WT、sun和ovate差异表达基因韦恩图分析，每一部分中黑色数字表示差异表达基因总数，分数的分子与分母分别表示上调与下调基因数，带有圆圈的字母表示该部分基因对应的热图；B：3个近等基因系共有差异表达基因中感兴趣基因热图；C ~ E：分别表示WT、sun和ovate中独有的差异表达基因中感兴趣基因热图；F：表示sun与ovate共有的差异表达基因中感兴趣基因热图；G：表示sun与WT共有的差异表达基因中感兴趣基因热图；H：表示ovate与WT共有的差异表达基因中感兴趣基因热图。

Fig. 4 Venn diagram and heatmap of the interested differentially expressed genes（DEGs）in the anthesis ovaries of WT，sun and ovate NILs after 2,4-D treatment A：Venn diagram of DEGs in WT，sun and ovate NILs after 2,4-D treatment. The black number in each part represents the total number of DEGs，and the numerator and denominator of the fraction represent the number of up-regulated and down-regulated genes，respectively. Letters with circles represent the heatmap corresponding to these genes. B：Heatmap of the interested DEGs shared by the 3 NILs；C-E：heatmaps of the interested genes that only differentially expressed in WT，sun and ovate NILs，respectively. F：Interested DEGs shared by sun and ovate. G：DEGs shared by sun and WT. H：DEGs shared by ovate and WT.

图5 2,4-D处理后各近等基因系花蕾差异表达基因的String分析 A：3个近等基因系共有差异表达基因分析；B：sun 独有的差异表达基因分析；C：ovate独有的差异表达基因分析；D：WT独有的差异表达基因分析；E：sun与ovate共有的差异表达基因分析；F：WT与ovate 共有的差异表达基因分析。

Fig. 5 String analysis of the differentially expressed genes（DEGs）caused by the 2,4-D treatment A：Analysis of DEGs that shared by the three NILs. B：Analysis of DEGs that were only in sun NIL. C：Analysis of DEGs that were only in ovate NIL. D：Analysis of DEGs that were only in WT NIL. E：Analysis of DEGs that were shared by sun and ovate NILs. F：Analysis of DEGs that were shared by WT and ovate NILs.

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