越橘FWL/PLAC8家族基因特征及表达分析

doi:10.16420/j.issn.0513-353x.2020-0151

摘要/Abstract

摘要：

以高丛越橘（Vaccinium corymbosum）大果型品种‘奥尼尔’和小果型品种‘蓝雨’为试材,测定花芽膨大与果实发育期间生长曲线及成熟果实的部分生理指标;开展包含PLAC8保守结构域的FWL（fruit weight like）基因全基因组鉴定与序列分析,并应用实时荧光定量PCR法检测VcFWL/PLAC8基因亚家族成员在花芽膨大与果实发育进程中的相对表达水平。结果表明：‘奥尼尔’果实单果质量与横径自S2期起均显著高于‘蓝雨’,其成熟果实中心室数、单果种子数及种子质量也显著高于‘蓝雨’果实。越橘VcFWL/PLAC8家族包含11个成员,分布于10条染色体上,结构分析显示这些基因均包含2 ~ 3个外显子。大部分VcFWL/PLAC8基因包含高度保守的结构域,聚类分析可将其分为3个亚家族,其中B和C亚家族成员间高度保守。qPCR分析表明A与C亚家族基因的表达模式和越橘花芽与果实发育进程中细胞增殖趋势一致,B亚家族基因的表达丰度较低,推测VcFWL/PLAC8基因可能通过不同的机制和途径参与调控果实的生长发育。

关键词: 越橘, VcFWL/PLAC8, 果实大小, 表达特性

Abstract:

Fruit size and weight are the most vital agronomic characters for consumers,breeders and researchers,and fruit weight（FW）was reported as an important quantitative trait locus affecting tomato and other fruit weight and size remarkably. In this study,the growth curves during Vaccinium corymbosum ‘O’Neal’（large-size fruit）and‘Bluerain’（small-size fruit）flower bud enlargement and fruit development,and conventionally physical indexes of mature fruits were measured and analyzed. The FWL family with conserved domain PLAC8 in the blueberry genome was identified,and the gene structure, chromosome location,conserved motifs of VcFWL/PLAC8 were analyzed. The relative expression level of VcFWL/PLAC8 genes during‘O’Neal’and‘Bluerain’flower bud enlargement and fruit development was detected by real-time fluorescence quantitative PCR. The results showed that the fruit weight of‘O’Neal’was dramatically higher than that of‘Bluerain’from stage S2,and the locule number,seed number and weight per fruit of‘O’Neal’mature fruits were also notably higher than those of‘Bluerain’. The VcFWL/PLAC8 family contained 11 members with highly conserved motifs in the‘Draper’genome,which can be divided into three clusters. qRT-PCR analysis showed that the expression patterns of A and C subfamilies were consistant with opposite to the cell proliferation trend,while the expression level of B subfamily was relatively low. These results suggested that VcFWL/PLAC8 genes might be involved in fruit growth and development through different regulatory mechanisms and pathways.

Key words: blueberry, FWL/PLAC8, Vaccinium corymbosum, fruit size, fruit development

中图分类号:

S663.9

俞蕾, 周雅, 宗宇, 张颖, 邱佳琪, 李永强, 杨莉, 郭卫东. 越橘FWL/PLAC8家族基因特征及表达分析[J]. 园艺学报, 2021, 48(2): 336-346.

YU Lei, ZHOU Ya, ZONG Yu, ZHANG Ying, QIU Jiaqi, LI Yongqiang, YANG Li, GUO Weidong. Characteristic and Relative Expression Pattern Analysis of FWL/PLAC8 Family in Blueberry[J]. Acta Horticulturae Sinica, 2021, 48(2): 336-346.

图/表 8

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引物名称 Primer name	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer	扩增长度/bp Predicted length
A subfamily	TCAGACATCCTTCCCACTAACC	CCGTCGCAGGAGACGTAG	138
B subfamily	CCAGGGTTTCGCTCTCCTTCC	AGCTCCCTTGGCCTGCGG	186
C subfamily	TCTGCGAAAGGTCTGGAGAT	CATGGCAACCAACATTTCAG	285
VcGAPDH	CGGCTACTTACGAGCAAATCAA	TTCAGTGTAGCCCAAAATTCCTTT	80

引物名称 Primer name	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer	扩增长度/bp Predicted length
A subfamily	TCAGACATCCTTCCCACTAACC	CCGTCGCAGGAGACGTAG	138
B subfamily	CCAGGGTTTCGCTCTCCTTCC	AGCTCCCTTGGCCTGCGG	186
C subfamily	TCTGCGAAAGGTCTGGAGAT	CATGGCAACCAACATTTCAG	285
VcGAPDH	CGGCTACTTACGAGCAAATCAA	TTCAGTGTAGCCCAAAATTCCTTT	80

品种 Cultivar	百粒种子质量/g Weight / 100 seeds	单果种子数 Seed number	单果种子质量/g Seed weight pre fruit	心室数比例/% Locule number
品种 Cultivar	百粒种子质量/g Weight / 100 seeds	单果种子数 Seed number	单果种子质量/g Seed weight pre fruit	4	5	6	7
奥尼尔O’Neal	0.04 ± 0.00	83.00 ± 4.06^**	0.036 ± 0.007^**		57.39	21.09	16.52
蓝雨Bluerain	0.04 ± 0.00	30.42 ± 8.31	0.012 ± 0.004	1.64	98.36

品种 Cultivar	百粒种子质量/g Weight / 100 seeds	单果种子数 Seed number	单果种子质量/g Seed weight pre fruit	心室数比例/% Locule number
品种 Cultivar	百粒种子质量/g Weight / 100 seeds	单果种子数 Seed number	单果种子质量/g Seed weight pre fruit	4	5	6	7
奥尼尔O’Neal	0.04 ± 0.00	83.00 ± 4.06^**	0.036 ± 0.007^**		57.39	21.09	16.52
蓝雨Bluerain	0.04 ± 0.00	30.42 ± 8.31	0.012 ± 0.004	1.64	98.36

基因名称 Gene name	染色体位置 Chromosome location	CDS/bp	外显子数 Exon number	大小/aa Size	分子量/kD Molecular weight	等电点 pI
gene-169.8	VaccDscaff5：16978476..16983996-	561	3	186	20.39	5.60
gene-244.19	VaccDscaff8：24434249..24439769+	693	2	230	25.72	4.92
gene-228.26	VaccDscaff10：22828768..22834442+	672	4	223	24.22	5.35
gene-180.23	VaccDscaff1：18011712..18017230-	777	3	258	27.84	8.12
gene-48.40	VaccDscaff27：4876429..4880014+	741	3	246	26.38	5.40
gene-85.17	VaccDscaff17：8569670..8572170+	657	2	218	23.42	8.54
gene-85.18	VaccDscaff17：8572400..8573369+	396	2	131	14.60	6.27
gene-305.30	VaccDscaff34：30578948..30582561-	738	3	246	26.40	5.37
gene-296.30	VaccDscaff29：29591240..29601268-	603	3	200	21.34	4.76
gene-308.41	VaccDscaff26：30828379..30838346-	603	3	200	21.34	4.76
gene-316.28	VaccDscaff21：31596223..31606319-	603	3	200	21.30	4.76
VcFW2.2	v.corymbosum_GDV_reftransV1_0004680	732	/	243	26.32	5.97