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作物学报 ›› 2019, Vol. 45 ›› Issue (3): 354-364.doi: 10.3724/SP.J.1006.2019.84095

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘蓝型油菜PEBP基因家族的鉴定与表达分析

荐红举,杨博,李阳阳,杨鸿,刘列钊,徐新福,李加纳()   

  1. 西南大学农学与生物科技学院 / 油菜工程研究中心 / 西南大学现代农业科学研究院, 重庆 400716
  • 收稿日期:2018-07-09 接受日期:2018-10-08 出版日期:2019-03-12 网络出版日期:2019-01-07
  • 通讯作者: 李加纳
  • 作者简介:荐红举, E-mail: jianhongju1989@126.com|杨博, E-mail: sheepneck@hotmail.com
  • 基金资助:
    本研究由高等学校学科创新引智基地项目“作物种质资源利用创新引智基地”(B12006);重庆市民生工程主题专项项目(cstc2016shms-ztzx80020);重庆市研究生创新项目资助(CYS17078)

Identification and expression analysis of PEBP gene family in oilseed rape

Hong-Ju JIAN,Bo YANG,Yang-Yang LI,Hong YANG,Lie-Zhao LIU,Xin-Fu XU,Jia-Na LI()   

  1. College of Agronomy and Biotechnology / Chongqing Engineering Research Center for Rapeseed / Academy of Agricultural Sciences, Southwest University, Chongqing 400715, China
  • Received:2018-07-09 Accepted:2018-10-08 Published:2019-03-12 Published online:2019-01-07
  • Contact: Jia-Na LI
  • Supported by:
    This study was supported by the Project of Intellectual Base for Discipline Innovation in Colleges and Universities(B12006);the Special Project of Chongqing People’s Livelihood(cstc2016shms-ztzx80020);Chongqing Graduate Student Research Innovation Project(CYS17078)

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摘要：

植物磷脂酰乙醇胺结合蛋白(phosphatidylethanolamine-binding protein, PEBP)基因对于控制开花时间具有重要作用。油菜作为世界上最重要的油料作物之一, 与模式植物拟南芥具有相似的开花习性。但有关油菜开花基因的功能研究相对较少。本研究利用拟南芥PEBP基因家族蛋白序列在油菜基因组内进行BlastP分析, 获得油菜PEBP家族成员, 并对其进行基因结构分析、motif预测、复制事件、进化树构建、选择压力分析和组织表达分析。结果表明, 共有26个油菜PEBP基因成员得到鉴定, 大部分成员含有4个外显子和3个内含子, motif-1和motif -2是PEBP成员的特征基序, 超过76.9%的成员属于片段复制扩增事件。进化树分析显示, PEBP分为3个亚家族, 基于转录组测序的组织表达数据表明油菜26个PEBP成员具有非常明显的组织表达特性。以上研究结果, 极大丰富了我们对甘蓝型油菜开花基因及调控模式的认识, 为进一步的分子育种提供了理论基础。

关键词: 油菜, 开花基因, PEBP, 进化, 表达

Abstract:

The plant phosphatidylethanolamine-binding protein (PEBP) genes play an important role in controlling flowering time. Rapeseed, as one of the most important oil crops in the world, has similar flowering habits with Arabidopsis thaliana. In this study, the PEBP family protein sequences of A. thaliana were used to perform BlastP analysis in the rapeseed genome. The members of the rape PEBP family were obtained, and the gene structure analysis, motif prediction, duplication analysis, phylogenetic tree construction, selective pressure analysis and tissue expression analysis were carried out on the family members. A total of 26 rapeseed PEBP gene members were identified, and most of them contained four exons and three introns, and motif-1 and motif-2 were the characteristics of PEBP members. Over 76.9% members were segmental duplicated. Phylogenetic tree analysis showed that PEBP was divided into three subfamilies. The tissue-specifics analysis based on RNA-Seq data showed that 26 PEBP members of rapeseed had very obvious tissue expression characteristics. All these results greatly enrich our understandings of flowering genes and regulation patterns in Brassica napus, and provide a theoretical basis for further molecular breeding in B. napus.

Key words: Brassica napus, flowering genes, PEBP, evolution analysis, expression analysis

附表1

‘中双11’的111个样品信息"

编号
Code		 生育期
Growth stage		 器官
Organ		 取样时间
Treatment time		 编号
Code		 生育期
Growth stage		 器官
Organ		 取样时间
Treatment time
Ro_24h 种子萌发 胚根 萌发后24 h Se_7d 青荚期 种子 花后7 d
Ro_48h 种子萌发 胚根 萌发后48 h Se_10d 青荚期 种子 花后10 d
Ro_72h 种子萌发 胚根 萌发后72 h Se_13d 青荚期 种子 花后13 d
Hy_24h 种子萌发 下胚轴 萌发后24 h Se_19d 青荚期 种子 花后19 d
Hy_48h 种子萌发 下胚轴 萌发后48 h Se_21d 青荚期 种子 花后21 d
Hy_72h 种子萌发 下胚轴 萌发后72 h Se_24d 青荚期 种子 花后24 d
Co_24h 种子萌发 子叶 萌发后24 h Se_27d 青荚期 种子 花后27 d
Co_48h 种子萌发 子叶 萌发后48 h Se_30d 青荚期 种子 花后30 d
Co_72h 种子萌发 子叶 萌发后72 h Se_35d 青荚期 种子 花后35 d
GS_12h 种子萌发 整个种子 萌发后12 h Se_40d 青荚期 种子 花后40 d
GS_24h 种子萌发 整个种子 萌发后24 h Se_43d 青荚期 种子 花后43 d
Ro_s 苗期-温室 根 Se_46d 青荚期 种子 花后46 d
Ro_s_f 苗期-大田 根 Se_49d 青荚期 种子 花后49 d
Ro_b 蕾期 根 Em_19d 青荚期 种胚 花后19 d
Ro_i 初花期 根 Em_21d 青荚期 种胚 花后21 d
Ro_f 盛花期 根 Em_24d 青荚期 种胚 花后24 d
St_b 蕾期 茎秆 Em_27d 青荚期 种胚 花后27 d
St_i 初花期 茎秆 Em_30d 青荚期 种胚 花后30 d
St_f 盛花期 茎秆 Em_35d 青荚期 种胚 花后35 d
St_24d 青荚期 茎秆 花后24 d Em_40d 成熟期 种胚 花后40 d
St_50d 成熟期 茎秆 花后50 d Em_43d 成熟期 种胚 花后43 d
Le_s 苗期-温室 叶片 Em_46d 成熟期 种胚 花后46 d
Le_s_f 苗期-大田 叶片 Em_49d 成熟期 种胚 花后49 d
LeY_b 蕾期 幼叶 Ra_40d 成熟期 胚芽 花后40 d
LeY_i 初花期 幼叶 Ra_43d 成熟期 胚芽 花后43 d
LeY_f 盛花期 幼叶 Ra_46d 成熟期 胚芽 花后46 d
LeY_10d 青荚期 幼叶 花后10 d Ra_49d 成熟期 胚芽 花后49 d
LeY_24d 青荚期 幼叶 花后24 d SC_19d 青荚期 种皮 花后19 d
LeY_30d 青荚期 幼叶 花后30 d SC_21d 青荚期 种皮 花后21 d
LeO_b 蕾期 成熟叶片 SC_27d 青荚期 种皮 花后27 d
LeO_i 初花期 成熟叶片 SC_30d 青荚期 种皮 花后30 d
LeO_f 盛花期 成熟叶片 SC_35d 青荚期 种皮 花后35 d
LeO_10d 青荚期 成熟叶片 花后10 d SC_40d 青荚期 种皮 花后40 d
LeO_24d 青荚期 成熟叶片 花后24 d SC_43d 成熟期 种皮 花后43 d
LeO_30d 青荚期 成熟叶片 花后30 d En_21d 青荚期 内种皮 花后21 d
Bu_b 蕾期 蕾 En_24d 青荚期 内种皮 花后24 d
Ao_i 初花期 花柄 Ep_24d 青荚期 外种皮 花后24 d
Ao_f 盛花期 花柄 Ep_30d 青荚期 外种皮 花后30 d
Cal_i 初花期 萼片 Fu_27d 青荚期 珠柄 花后27 d
Cal_f 盛花期 萼片 Fu_35d 青荚期 珠柄 花后35 d
Pe_i 初花期 花瓣 SP_3d 青荚期 角果皮 花后3 d
Pe_f 盛花期 花瓣 SP_5d 青荚期 角果皮 花后3 d
Pi_f_un 盛花期 雌蕊-未授粉 SP_7d 青荚期 角果皮 花后7 d
Pi_i 初花期 雌蕊 SP_10d 青荚期 角果皮 花后10 d
Pi_f 盛花期 雌蕊 SP_13d 青荚期 角果皮 花后13 d
Sta_i 初花期 雄蕊 SP_16d 青荚期 角果皮
Sta_f 盛花期 雄蕊 SP_19d 青荚期 角果皮 花后19 d
At_i 初花期 花药 SP_21d 青荚期 角果皮 花后21 d
At_f 盛花期 花药 SP_24d 青荚期 角果皮 花后24 d
Cap_i 初花期 花丝 SP_27d 青荚期 角果皮 花后27 d
Cap_f 盛花期 花丝 SP_30d 青荚期 角果皮 花后30 d
IT_b 蕾期 主序顶端 SP_35d 青荚期 角果皮 花后35 d
IT_i 初花期 主序顶端 SP_40d 成熟期 角果皮 花后40 d
IT_f 盛花期 主序顶端 SP_43d 成熟期 角果皮 花后43 d
Se_3d 青荚期 种子 花后3 d SP_46d 成熟期 角果皮 花后46 d
Se_5d 青荚期 种子 花后3 d

表1

利用BlastP方法获得油菜、白菜、甘蓝和拟南芥PEBP同源基因"

AGI ID 油菜同源基因
B. napus homologs		 白菜或甘蓝同源基因
B. rapa or B. oleracea homologs		 基因模型
Gene models
AT5G62040 BnaA06g21490D Bra010052 BROTHER OF FT AND TFL1 (BFT)
AT5G62040 BnaC03g52010D Bol012573 BROTHER OF FT AND TFL1 (BFT)
AT5G62040 -— Bol015337 BROTHER OF FT AND TFL1 (BFT)
AT2G27550 BnaA03g22340D Bra000506 centroradialis (ATC)
AT2G27550 BnaA04g15930D Bra034357 centroradialis (ATC)
AT2G27550 BnaA07g13290D Bra012010 centroradialis (ATC)
AT2G27550 BnaC01g41050D -— centroradialis (ATC)
AT2G27550 BnaC03g47080D Bol026421 centroradialis (ATC)
AT2G27550 BnaC03g64560D -— centroradialis (ATC)
AT2G27550 BnaC04g39220D Bol032920 centroradialis (ATC)
AT2G27550 BnaC04g16750D -— centroradialis (ATC)
AT1G65480 BnaA02g12130D Bra022475 FLOWERING LOCUS T (FT)
AT1G65480 -— Bra004117 FLOWERING LOCUS T (FT)
AT1G65480 BnaC02g45250D Bol017639 FLOWERING LOCUS T (FT)
AT1G65480 BnaC04g14850D Bol039209 FLOWERING LOCUS T (FT)
AT1G18100 BnaA09g44700D Bra031016 MOTHER OF FT AND TFL1 (MFT)
AT1G18100 BnaA06g12320D Bra025930 MOTHER OF FT AND TFL1 (MFT)
AT1G18100 BnaC05g50120D Bol009744 MOTHER OF FT AND TFL1 (MFT)
AT1G18100 BnaC08g37400D Bol030802 MOTHER OF FT AND TFL1 (MFT)
AT5G03840 BnaA10g26300D Bra009508 TERMINAL FLOWER 1 (TFL1)
AT5G03840 BnaAnng00810D Bra028815 TERMINAL FLOWER 1 (TFL1)
AT5G03840 -— Bra005783 TERMINAL FLOWER 1 (TFL1)
AT5G03840 BnaC02g02900D Bol005471 TERMINAL FLOWER 1 (TFL1)
AT5G03840 BnaC03g01440D Bol010027 TERMINAL FLOWER 1 (TFL1)
AT5G03840 BnaCnng10680D Bol015337 TERMINAL FLOWER 1 (TFL1)
AT4G20370 BnaA07g25310D Bra022475 TWIN SISTER OF FT (TSF)
AT4G20370 BnaA07g33120D Bra015710 TWIN SISTER OF FT (TSF)
AT4G20370 BnaC02g23820D -— TWIN SISTER OF FT (TSF)
AT4G20370 BnaC06g27090D -— TWIN SISTER OF FT (TSF)

图1

油菜PEBP进化树与基因结构"

图2

油菜PEBP基因家族motif预测"

图3

油菜与拟南芥PEBP蛋白序列多重比对"

图4

油菜、白菜、甘蓝和拟南芥PEBP基因家族进化分析"

表2

油菜和拟南芥PEBP同源基因的Ka/Ks"

油菜基因编号
Gene ID in B. napus		 拟南芥基因编号
Gene ID in A. thaliana		 基因模型
Gene model		 非同义突变频率
Ka		 同义突变频率
Ks		 Ka/Ks
BnaA02g12130D AT1G65480 FT 0.09 0.32 0.27
BnaC02g45250D AT1G65480 FT 0.08 0.35 0.24
BnaC04g14850D AT1G65480 FT 0.11 0.80 0.14
BnaA10g26300D AT5G03840 TFL1 0.08 0.37 0.23
BnaAnng00810D AT5G03840 TFL1 0.05 0.39 0.14
BnaC02g02900D AT5G03840 TFL1 0.05 0.39 0.14
BnaC03g01440D AT5G03840 TFL1 0.03 0.41 0.07
BnaCnng10680D AT5G03840 TFL1 0.08 0.35 0.22
BnaA07g25310D AT4G20370 TSF 0.10 0.80 0.12
BnaA07g33120D AT4G20370 TSF 0.09 0.45 0.21
BnaC02g23820D AT4G20370 TSF 0.13 0.45 0.30
BnaC06g27090D AT4G20370 TSF 0.23 0.88 0.26
BnaA03g22340D AT2G27550 ATC 0.02 0.31 0.07
BnaA04g15930D AT2G27550 ATC 0.03 0.33 0.08
BnaA07g13290D AT2G27550 ATC 0.03 0.49 0.06
BnaC01g41050D AT2G27550 ATC 0.03 0.33 0.08
BnaC03g47080D AT2G27550 ATC 0.03 0.31 0.10
BnaC03g64560D AT2G27550 ATC 0.03 0.33 0.08
BnaC04g16750D AT2G27550 ATC 0.03 0.52 0.07
BnaC04g39220D AT2G27550 ATC 0.06 0.39 0.15
BnaA06g21490D AT5G62040 BFT 0.05 0.31 0.17
BnaC03g52010D AT5G62040 BFT 0.05 0.31 0.18
BnaA09g44700D AT1G18100 MFT 0.02 0.40 0.05
BnaC08g37400D AT1G18100 MFT 0.02 0.45 0.05
BnaC05g50120D AT1G18100 MFT 0.03 0.37 0.08
BnaA06g12320D AT1G18100 MFT 0.03 0.37 0.09

图5

油菜PEBP家族成员的组织表达分析 横坐标样品名称详见附表1, 蓝色表示低表达, 黄色表示高表达。"

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