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Molecular Cloning and Expression Analysis of the ζ-Carotene Desaturase Gene in Chinese kale(Brassica oleracea var.alboglabra Bailey) 被引量:10
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作者 SUN Bo ZHANG Fen +6 位作者 XUE Shengling CHANG Jiaqi ZHENG Aihong JIANG Min MIAO Huiying WANG Qiaomei TANG Haoru 《Horticultural Plant Journal》 SCIE 2018年第3期94-102,共9页
ζ-Carotene desaturase(ZDS)is an important enzyme in carotenoid biosynthesis.Here,the Brassica oleracea var.alboglabra ZDS(Boa ZDS)gene was cloned from Chinese kale via reverse transcription-polymerase chain reaction(... ζ-Carotene desaturase(ZDS)is an important enzyme in carotenoid biosynthesis.Here,the Brassica oleracea var.alboglabra ZDS(Boa ZDS)gene was cloned from Chinese kale via reverse transcription-polymerase chain reaction(RT-PCR)and deposited in Gen Bank(accession number KY662297).The Boa ZDS gene contains an open reading frame of 1 686 bp that encodes a 561-amino acid protein.Sequence analysis indicates that the ZDS protein is apparently conserved during plant evolution and is most closely related to B.oleracea var.capitata and B.rapa.The promoter sequence of the Boa ZDS gene was predicted to harbor several cis-acting elements that are related to light and phytohormone responses.Semiquantitative RT-PCR analysis showed that Boa ZDS expression varied among different developmental stages and organs.Relative ZDS expression remained stable during germination and seedling stages and rapidly increased at the mature leaf stage.The leaves showed the highest ZDS expression levels compared to the other organs.ZDS expression decreased in all flower tissues during blooming.The fused protein of Boa ZDS was obtained by prokaryotic expression.Heterologous expression of Boa ZDS in Escherichia coli confirmed that Boa ZDS encodes a functionalζ-carotene desaturase that increases β-carotene accumulation in E.coli cells harboring a β-carotene-producing plasmid.The findings of the present study provide a molecular basis for the elucidation of ZDS gene function in Chinese kale. 展开更多
关键词 Chinese kale brassica oleracea var alboglabra Bailey carotenoids ζ-carotene desaturase(ZDS) CLONING gene expression prokaryotic expression heterologous expression
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花椰菜(Brassica oleracea var.botrytis)黑腐病抗性基因同源序列分离及克隆的研究 被引量:3
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作者 古瑜 赵前程 +3 位作者 刘松 王春国 孙德岭 宋文芹 《南开大学学报(自然科学版)》 CAS CSCD 北大核心 2007年第2期62-66,共5页
通过从 NBS 保守序列设计简并引物 PCR 的方法,以花椰菜(Brassica oleracea vat.botrytis)抗、感黑腐病的近等基因系为材料,分离得到 NBS-LRR 型抗性基因同源序列,并获得1个克隆,命名为 RGA330-7.Southern 杂交表明,该片段在近等基因系... 通过从 NBS 保守序列设计简并引物 PCR 的方法,以花椰菜(Brassica oleracea vat.botrytis)抗、感黑腐病的近等基因系为材料,分离得到 NBS-LRR 型抗性基因同源序列,并获得1个克隆,命名为 RGA330-7.Southern 杂交表明,该片段在近等基因系中存在明显的多态性,且该片段在抗黑腐病基因位点至少存在3个以上类似 RGA330-7的同源拷贝.序列分析结果认为该克隆与 NBS-LRR 型抗性基因的部分 CDSs 有很高的同源性,说明该片段属于 NBS-LRR 型.系统进化分析该序列与甘蓝型油菜的2个抗病同源序列归为一类,很可能这3个不同来源的抗性基因同源序列同属于一种抗性基因家族.因此推测该序列与花椰菜抗黑腐病基因紧密相关,为进一步克隆花椰菜抗黑腐病基因提供了可靠的候选基因,对分子标记辅助抗性育种具有重要意义。 展开更多
关键词 黑腐病 花椰菜(brassica oleracea var.botrytis) RGAs(resistance gene analogs RGAs)
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花椰菜(Brassica oleracea var.botrytis)抗黑腐病差异表达cDNA片段的克隆及功能的初步研究 被引量:5
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作者 古瑜 毛英伟 +3 位作者 赵前程 孙德岭 刘惠静 宋文芹 《南开大学学报(自然科学版)》 CAS CSCD 北大核心 2008年第4期42-48,共7页
采用花椰菜抗黑腐病近等基因系C731(感病系)和C712(抗病系)作为材料,利用cDNA—AFLP技术,研究了花椰菜黑腐病抗性在黑腐病菌侵染和非侵染条件下基因表达的情况.获得了两个阳性克隆M2和M6.Northern杂交和点杂交进一步证明M2是组... 采用花椰菜抗黑腐病近等基因系C731(感病系)和C712(抗病系)作为材料,利用cDNA—AFLP技术,研究了花椰菜黑腐病抗性在黑腐病菌侵染和非侵染条件下基因表达的情况.获得了两个阳性克隆M2和M6.Northern杂交和点杂交进一步证明M2是组成型表达的cDNA片段,只是在病菌侵染与非侵染条件下表达丰度不同.而M6是差异表达的cDNA片段.Southern杂交表明M6 cDNA片段在花椰菜基因组中是单拷贝序列.随后的序列分析发现M6cDNA片段与拟南芥1号染色体的BACF19P19中66665~66813bp有84%同源性,该片段编码拟南芥的2A6蛋白的部分序列.推测的氨基酸序列与拟南芥的2A6蛋白部分序列有91%同源性.用H2O2作为外源分子胁迫处理的初步功能分析发现:M6cDNA片段受H2O2诱导,在诱导早期16~24h高度表达,其在叶片中的积累呈逐渐上升趋势.根据序列分析和初步的功能分析的结果推测:M6cDNA片段可能就是编码花椰菜中类2A6蛋白的部分基因片段.是参与花椰菜抗病反应信号途径的相关调控基因片段. 展开更多
关键词 花椰菜(brassica oleracea VaF.botrytis) 黑腐病 抗病相关基因片段 cDNA—AFLP
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Molecular Cloning and Sequence Analysis ofBoPGIP2 Gene from Brassica oleracea L. var. alboglabra
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作者 张弢 《Agricultural Science & Technology》 CAS 2009年第4期91-95,104,共6页
PGIP gene was obtained from Brassica oleracea L. var. alboglabra, named BoPGIP2. The full length of BoPGIP2 gene is 1 102 bp and the exon is 993 bp which encodes a protein of 330 amino acids with a predicted molecular... PGIP gene was obtained from Brassica oleracea L. var. alboglabra, named BoPGIP2. The full length of BoPGIP2 gene is 1 102 bp and the exon is 993 bp which encodes a protein of 330 amino acids with a predicted molecular mass of 37.1 kDa, interrupted by one intron of 95 bp in, length. Sequence analysis revealed that it has five potential N-giycosylation sites, two protein kinase C phosphrylation sites, five casin kinase Ⅱ phosphrylation sites and four N-myristoylation sites. The amino acids sequences alignment confirmed that ^145 LRR stucture was highly conserved in all aligned PGIP sequences. 展开更多
关键词 brassica oleracea L. vat. alboglabra BoPGIP2 Molecular cloning Sequence analysis
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花椰菜(Brassica oleracea var.botrytis L.)凝集素的细胞凝集和糖抑制作用的研究 被引量:10
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作者 黄德棋 余萍 +2 位作者 朱苏闽 颜志森 林玉满 《福建师范大学学报(自然科学版)》 CAS CSCD 1992年第2期72-77,共6页
花椰莱(Brassica oleracea var. botrytis L.)凝集素能凝集兔、大鼠和小鼠的红细胞,其中对兔的红细胞的凝集活性最高,最低凝集浓度为0.68μg/ml。但不凝集我们所测试的其它16种红细胞。浓度为0.1mg/ml时,花椰菜凝集素也能凝集小鼠艾氏... 花椰莱(Brassica oleracea var. botrytis L.)凝集素能凝集兔、大鼠和小鼠的红细胞,其中对兔的红细胞的凝集活性最高,最低凝集浓度为0.68μg/ml。但不凝集我们所测试的其它16种红细胞。浓度为0.1mg/ml时,花椰菜凝集素也能凝集小鼠艾氏腹水瘤细胞,S_(180)肉瘤细胞,大鼠W_(256)肿瘤细胞、人的MGC_(80-3)胃癌细胞、小鼠和大鼠的脾脏淋巴细胞、骨髓细胞以及牛精子细胞,而不凝集人的Hela细胞。该凝集素对免的红细胞的凝集活性可被L—鼠李糖和D—树胶醛糖所抑制,最低抑制浓度分别为33.3m mol/L和16.7m mol/L。 展开更多
关键词 花椰菜 凝集素 糖抑制 细胞凝集
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Transformation of insect-resistant gene into cauliflower (Brassica oleracea L.var. botrytis)
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作者 吕玲玲 雷建军 +2 位作者 宋明 李立云 曹必好 《Agricultural Science & Technology》 CAS 2004年第3期17-21,共5页
Cowpea Trypsin Inhibitor (CpTI) gene was transferred into the cotyle dons and hypocotyls of cauliflower by Agrobacterium-mediated transformation met hod. The best selective concentration of kanamycin (kan) was 15 mg L... Cowpea Trypsin Inhibitor (CpTI) gene was transferred into the cotyle dons and hypocotyls of cauliflower by Agrobacterium-mediated transformation met hod. The best selective concentration of kanamycin (kan) was 15 mg L-1. The con centration of carbencillin (carb) was 500 mg L-1. 14 transgenic cauliflower pla nts were obtained. The putative transformants were assayed by PCR and Southern b lotting analysis. The results indicated that CpTI gene was transferred into caul iflower successfully. 展开更多
关键词 cauliflower (brassica oleracea L. var. botrytis) Agrobacterium-me diated genetic transformation CpTI gene transgenic plant
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A Co-Dominant Marker BoE332 Applied to Marker-Assisted Selection of Homozygous Male-Sterile Plants in Cabbage (Brassica oleracea var.capitata L.) 被引量:1
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作者 CHEN Chen ZHUANG Mu +5 位作者 FANG Zhi-yuan WANG Qing-biao ZHANG Yang-yong LIU Yu-mei YANG Li-mei CHENG Fei 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2013年第4期596-602,共7页
The dominant genic male sterility (DGMS) gene CDMs399-3 derived from a spontaneous mutation in the line 79-399-3 of spring cabbage (Brassica oleracea var. capitata L.), has been successfully applied in hybrid seed... The dominant genic male sterility (DGMS) gene CDMs399-3 derived from a spontaneous mutation in the line 79-399-3 of spring cabbage (Brassica oleracea var. capitata L.), has been successfully applied in hybrid seed production of several cabbage cultivars in China. During the development of dominant male sterility lines in cabbage, the conventional identification of homozygous male-sterile plants (CDMs399-3/CDMs399-3) is a laborious and time-consuming process. For marker-assisted selection (MAS) of the gene CDMs399-3 transferred into key spring cabbage line 397, expressed sequence tag-simple sequence repeats (EST-SSR) and SSR technology were used to identify markers that were linked to CDMs399-3 based on method of bulked segregant analysis (BSA). By screening a set of 978 EST-SSRs and 395 SSRs, a marker BoE332 linked to the CDMs399-3 at a distance of 3.6 cM in the genetic background of cabbage line 397 were identified. 7 homozygons male-sterile plants in population P1170 with 20 plants were obtained finally via MAS of BoE332. Thus, BoE332 will greatly facilitate the transferring of the gene CDMs399-3 into the key spring cabbage line 397 and improve the application of DGMS in cabbage hybrid breeding. 展开更多
关键词 cabbage brassica oleracea var. capitata L.) dominant genic male sterility (DGMS) expressed sequencetag-simple sequence repeats (EST-SSR) bulked segregant analysis (BSA) marker-assisted selection (MAS)
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The protoplasts isolation,culture and shoots regeneration of broccoli(Brassica oleracea var.italica)
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作者 王怀名 A.Schfer-Menuhr G.Mix-Wagner 《华北农学报》 CSCD 北大核心 1993年第S2期28-34,共7页
Eight F<sub>1</sub>-hybrid cultivars of broccoli were studied.We obtained cell division,celled colonies and p-calli in 5 cultivars,roots and shoots regeneration in one cultivar.The leavesof propagated plan... Eight F<sub>1</sub>-hybrid cultivars of broccoli were studied.We obtained cell division,celled colonies and p-calli in 5 cultivars,roots and shoots regeneration in one cultivar.The leavesof propagated plantlets in vitro were cut into 1—2mm pieces,isolated with an enzyme solutioncontaining 2% cellulase and 1%macerase on a rotary shaker(50 rpm,21℃,3h,2500 lux light),and purified with a 0.5M sucrose solution.The purified protoplasts were placed on a drop of 1%agarose.2—3 ml liquid medium was added around the agarose drops,and all of the cultures wereincubated at 25℃ under light(4000 lux)for 16 hours.3—5 days after isolation the cell divisionwas found.About 7 days after incubation 4 multicellular colonies were formed.After 3—5 wksome p-calli were developed.When the p-calli were 2—3 mm in diameter it was transferred to asolidified medium.Once they were developed to 1 cm in diameter they were transferred on a re-generation medium.About 5 months after incubation some roots and shoots grown from the calliwere 展开更多
关键词 PROTOPLASTS CULTURE SHOOTS REGENERATION broccoli( brassica oleracea var. ITALICA )
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Review of Research Situation of Brassica oleracea var.italica
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作者 Ying GAO 《Asian Agricultural Research》 2020年第7期61-63,共3页
Broccoli(Brassica oleracea var.italica),also called green broccoli,green cauliflower,and calabrese,belongs to genus Brassica in family Cruciferae,and is annual or biennial herbaceous plant.Both broccoli and cauliflowe... Broccoli(Brassica oleracea var.italica),also called green broccoli,green cauliflower,and calabrese,belongs to genus Brassica in family Cruciferae,and is annual or biennial herbaceous plant.Both broccoli and cauliflower are varieties of Brassica oleracea L.Broccoli is rich in nutrients.Its protein,amino acids and vitamins are higher than cauliflower,and broccoli is easy to grow,the supply period is long,and it has a good market and economic value.This paper introduced broccoli-related information from broccoli's nutritional value,cultivation techniques,existing problems and prospects.In addition,on the basis of existing studies,it discussed the future development prospects of broccoli,in order to promote the production research of broccoli in China. 展开更多
关键词 Broccoli(brassica oleracea var.italica) Nutritional value Cultivation techniques Field management PROSPECTS
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Screening and Identifying Two Specifi c Molecular Markers in Maintainer Line of Cytoplasmic Male Sterility Cauliflower (Brassica oleracea var. botrytis)
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作者 C.G. Wang  J.M. Gao  +3 位作者 H. Li  Y. Gu  X.Q. Chen  W.Q. Song 《分子植物育种》 CAS CSCD 2007年第2期269-270,共2页
Cytoplasmic male sterility (CMS) is a maternally inherited trait that prevents the production of function pollen, but maintains female fertility. It has been widely used in breeding programs to product F_1 hybrid seed
关键词 分子标记 花椰菜 细胞质 种植技术
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流式细胞仪对甘蓝花粉倍性的测定
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作者 杨鼎 李崇娟 +4 位作者 吕凤仙 和江明 兰梅 胡靖峰 徐学忠 《湖北农业科学》 2024年第4期78-81,117,共5页
利用流式细胞仪测定甘蓝(Brassica oleracea var.capitata)花粉的倍性,以甘蓝GL21028的花粉为材料,以叶片为对照,比较花粉2种处理方式、3种不同细胞裂解液、2种核酸染料的效果差异。通过B缓冲液处理后获取的花粉细胞可以直接染色,为最... 利用流式细胞仪测定甘蓝(Brassica oleracea var.capitata)花粉的倍性,以甘蓝GL21028的花粉为材料,以叶片为对照,比较花粉2种处理方式、3种不同细胞裂解液、2种核酸染料的效果差异。通过B缓冲液处理后获取的花粉细胞可以直接染色,为最简便处理方式;采用“Aru”buffer和Y 2种裂解液,染色剂DAPI,处理甘蓝叶片和花粉,通过流式细胞仪检测,样本细胞DNA成峰集中,细胞碎片很少;配制的Y裂解液能够适用甘蓝材料流式细胞仪的观察。甘蓝花粉通过B缓冲液处理后,可以直接作为流式细胞仪倍性鉴定的材料,不再需要其他处理,配制的Y裂解液可以得到较好的试验结果。 展开更多
关键词 甘蓝(brassica oleracea var.capitata) 流式细胞仪 花粉 测定 倍性 裂解液
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BoMyrosinase plays an essential role in sulforaphane accumulation in response to selenite treatment in broccoli
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作者 Qiuyun Wu Qi Wu +4 位作者 Yuxiao Tian Chunyan Zhou Shuxiang Mao Junwei Wang Ke Huang 《Horticultural Plant Journal》 SCIE CAS CSCD 2024年第2期488-502,共15页
Sulforaphane, a naturally specialized metabolite, plays significant roles in human disease prevention and plant defense. Myrosinase(MY) is a key gene responsible for the catalysis of sulforaphane formation, but the mo... Sulforaphane, a naturally specialized metabolite, plays significant roles in human disease prevention and plant defense. Myrosinase(MY) is a key gene responsible for the catalysis of sulforaphane formation, but the molecular mechanisms through which MY regulates sulforaphane biosynthesis in plants remains largely unknown. Here, we discovered that the change of sulforaphane content in broccoli sprouts caused by exogenous selenite treatments is positively related to BoMY expression. BoMY overexpression in the Arabidopsis thaliana tgg1 mutants could dramatically increase myrosinase activity and sulforaphane content in the rosette leaves of 35S::BoMY/tgg1 and rescue its phenotypes.Moreover, an obvious increase of myrosinase activity and sulforaphane content was displayed in transgenic BoMY-overexpressed broccoli lines.In addition, a 2 033 bp promoter fragment of BoMY was isolated. Yeast one-hybrid(Y1H) library screening experiment uncovered that one bHLH transcription factor, BoFAMA, could directly bind to BoMY promoter to activate its expression, which was further evidenced by Y1H assay and dual-luciferase reporter assay. BoFAMA is a selenite-responsive transcription factor that is highly expressed in broccoli leaves;its protein is solely localized to nucleus. Additionally, genetic evidence suggested that the knockdown of FAMA gene in Arabidopsis thaliana could significantly decrease sulforaphane yield by inhibiting the expression of myrosinase genes. Interestingly, exogenous selenite supply could partially restore the low level of sulforaphane content in transgenic Arabidopsis FAMA-silencing plants. Our findings uncover a novel function of FAMAMY module in the regulation of selenite-mediated sulforaphane synthesis and provide a new insights into the molecular mechanism by which selenite regulates the accumulation of sulforaphane in plants. 展开更多
关键词 brassica oleracea var.italica MYROSINASE BoMY SULFORAPHANE Selenite treatment
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Comparison of carotenoid,chlorophyll concentrations and their biosynthetic transcript levels in different coloured cauliflower
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作者 Fatemeh Izadpanah Katja Frede +1 位作者 Forouzandeh Soltani Susanne Baldermann 《Horticultural Plant Journal》 SCIE CAS CSCD 2024年第3期743-754,共12页
Carotenoids and chlorophylls are among the most widely distributed pigments in nature that play essential roles in the photosynthetic apparatus and confer diverse colours in plants.Among all vegetables,cauliflower(Bra... Carotenoids and chlorophylls are among the most widely distributed pigments in nature that play essential roles in the photosynthetic apparatus and confer diverse colours in plants.Among all vegetables,cauliflower(Brassica oleracea L.ssp.var.botrytis)is rich in phytochemicals and is an important crop grown all over the world.This study investigates carotenoid and chlorophyll concentrations in differently pigmented cultivars and elucidates the role of transcriptional regulation of carotenoid accumulation including lutein andβ-carotene.Here,we characterised changes in pigments by UHPLC-DAD-ToF-MS and changes in transcript levels of carotenoid metabolic genes by qRT-PCR in florets and leaves of orange(‘Jaffa'and‘Sunset'),purple(‘Di Sicilia Violetto'and‘Graffiti'),green(‘Trevi')and white(‘Clapton')cultivars.Transcript levels of all carotenoid metabolic genes showed different transcript level patterns in the leaves and florets.Compared to the other cultivars,the orange cultivars had the highest levels ofβ-carotene in the florets and lutein in the leaves resulting in changes lutein/β-carotene ratios.In the green cultivar,higher transcript levels were also found,especially for phytoene synthase and phytoene desaturase genes of the core biosynthesis pathway.However,no increased carotenoid concentrations were observed,possibly due to a higher carotenoid turnover induced by the carotenoid cleavage dioxygenase 4 in the green cultivar.In the white(‘Clapton')and purple(‘Di Sicilia Violetto'and‘Graffiti')cultivars the phytoene desaturase transcript levels as well as carotenoid concentrations were low.Chlorophyll concentrations changed in trend comparable to the carotenoid concentrations and were only significantly lower in the leaves of the orange cultivar‘Jaffa'.Also,the chlorophyll a/b ratio changed in‘Jaffa'.In florets the highest chlorophylls concentrations were observed for the green cultivar(‘Trevi')and the purple cultivar(‘Di Sicilia Violetto').Taken together,the study demonstrates the complex source-sink relationship of carotenoid accumulation in different coloured cauliflower. 展开更多
关键词 brassica oleracea L.var.botrytis CAROTENOID LUTEIN Β-CAROTENE BIOSYNTHESIS qRT-PCR
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Expression changes of mi RNA-regulated genes associated with the formation of the leafy head in cabbage
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作者 Jorge Aleman-Baez Jose Fernando Acevedo-Zamora +3 位作者 Johan Bucher Chengcheng Cai Roeland E.Voorrips Guusje Bonnema 《Horticultural Plant Journal》 SCIE CAS CSCD 2024年第4期1007-1019,共13页
The vegetative development of cabbage(Brassica oleracea var.capitata)passes through seedling,rosette,folding and heading stages.Leaves that form the rosette are large and mostly flat.In the following developmental sta... The vegetative development of cabbage(Brassica oleracea var.capitata)passes through seedling,rosette,folding and heading stages.Leaves that form the rosette are large and mostly flat.In the following developmental stages,the plants produce leaves that curve inward to produce the leafy head.Many microRNAs and their target genes have been described participating in leaf development and leaf curvature.The aim of this study is to investigate the role of miRNA-regulated genes in the transition from the rosette to the heading stage.We compared the mi RNA and gene abundances between emerging rosette and heading leaves.To remove transcripts(miRNAs and genes)whose regulation was most likely associated with plant age rather than the change from rosette to heading stage,we utilized a non-heading collard green(B.oleracea var.acephala)morphotype as control.This resulted in 33 DEMs and 1998 DEGs with likely roles in the transition from rosette to heading stage in cabbage.Among these 1998 DEGs,we found enriched GO terms related to DNA-binding transcription factor activity,transcription regulator activity,iron ion binding,and photosynthesis.We predicted the target genes of these 33 DEMs and focused on the subset that was differentially expressed(1998DEGs)between rosette and heading stage leaves to construct mi RNA-target gene interaction networks.Our main finding is a role for miR396b-5p targeting two Arabidopsis thaliana orthologues of GROWTH REGULATING FACTORs 3(GRF3)and 4(GRF4)in pointed cabbage head formation. 展开更多
关键词 brassica oleracea var.capitata RNA sequencing miRNA-target gene network leafy head formation
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Effect of H_2O_2 on Growth of Collard( Brassis oleracea L. ) Seedlings Under Salt Stress
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作者 Wei LI Junjie GUO Hongyan LI 《Agricultural Biotechnology》 CAS 2019年第1期43-46,184,共5页
Collard variety( Brassis oleracea L. var. acephala f. tricolor Hort.) as a research material was treated with exogenous H_2O_2 and H_2O_2 scavenger dimethyl thiourea under 100 mmol/L NaC l stress. Two days later,growt... Collard variety( Brassis oleracea L. var. acephala f. tricolor Hort.) as a research material was treated with exogenous H_2O_2 and H_2O_2 scavenger dimethyl thiourea under 100 mmol/L NaC l stress. Two days later,growth rate,dry weight,fresh weight and relative water content of the plants were determined. After 6h of treatment,the activity and gene expression of three antioxidant enzymes,superoxide dismutase( SOD),catalase( CAT) and ascorbate peroxidase( APX) in plants,were measured. The results showed that the growth rate,dry weight,fresh weight,relative water content,and the activity and gene expression of the three antioxidant enzymes in collard seedlings were higher in the treatment of salt stress with the addition of 0. 05 mmol/L exogenous H_2O_2 than in the simple salt stress treatment; and when endogenous H_2O_2 was removed,the growth rate,dry weight,fresh weight,relative water content,and the activity and gene expression of the three antioxidant enzymes in plant seedlings were lower than those under simple salt stress. It is speculated that under salt stress,H_2O_2 is involved in the regulation of antioxidant defense gene expression,and it might be an important regulator of salt-induced antioxidant system in collard leaves. 展开更多
关键词 brassica oleracea L.var.acephala f.tricolor Hort. H 2O 2 Salt stress Antioxidase Gene expression
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青花菜雄性不育相关基因BoDHAR的克隆与表达分析 被引量:7
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作者 张国裕 康俊根 +2 位作者 张延国 程智慧 王晓武 《生物工程学报》 CAS CSCD 北大核心 2006年第5期751-756,共6页
以一个与甘蓝显性核不育相关的差异表达片段的序列为信息探针,通过在NCBI与TAIR网站数据库中进行同源EST序列搜索,经人工拼接、RT-PCR、PCR克隆与序列分析,获得了青花菜脱氢抗坏血酸还原酶DHARdehydroascorbatereductase基因的cDNA与DN... 以一个与甘蓝显性核不育相关的差异表达片段的序列为信息探针,通过在NCBI与TAIR网站数据库中进行同源EST序列搜索,经人工拼接、RT-PCR、PCR克隆与序列分析,获得了青花菜脱氢抗坏血酸还原酶DHARdehydroascorbatereductase基因的cDNA与DNA全长序列,命名为BoDHAR。并利用双链接头介导PCR的染色体步行技术(genomewalking)克隆了其上游644bp的5′端序列。所获的BoDHAR基因全长1486bp,存在两个内含子,DNA编码区序列633bp,编码210个氨基酸;序列分析表明BoDHAR与同源基因AT1G19570.1cDNA序列有82.3%的一致性,推导的氨基酸序列有79.6%的一致性;编码的水溶性蛋白存在多个磷酸化位点;5′端上游区存在明显的转录调控序列。半定量RT-PCR结果表明BoDHAR在可育系花蕾中的表达量明显高于不育系花蕾,在花药中的表达明显高于其它部位。 展开更多
关键词 青花菜(brassica oleracea L. var. ITALICA ) DHAR(dehydroascorbate reductase) 基因克隆 RT-PCR
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甘蓝类蔬菜小孢子再生植株染色体倍性与气孔保卫细胞叶绿体数的相关性 被引量:22
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作者 袁素霞 刘玉梅 +4 位作者 方智远 杨丽梅 庄木 张扬勇 孙培田 《中国农业科学》 CAS CSCD 北大核心 2009年第1期189-197,共9页
【目的】研究甘蓝类蔬菜游离小孢子再生植株的染色体倍性与气孔保卫细胞叶绿体数的相关性,为甘蓝类蔬菜提供一种快速、简易、经济、实用而又可靠的倍性鉴定方法。【方法】采用分布型分析和t测验,对结球甘蓝、青花菜和芥蓝不同倍性的小... 【目的】研究甘蓝类蔬菜游离小孢子再生植株的染色体倍性与气孔保卫细胞叶绿体数的相关性,为甘蓝类蔬菜提供一种快速、简易、经济、实用而又可靠的倍性鉴定方法。【方法】采用分布型分析和t测验,对结球甘蓝、青花菜和芥蓝不同倍性的小孢子再生植株的气孔保卫细胞叶绿体数进行统计分析。以根尖染色体计数法和田间形态学观察法的倍性鉴定结果,对叶绿体数分界法的可靠性进行验证。【结果】同一倍性植株上的不同叶片间及同一叶片的不同部位间,气孔保卫细胞叶绿体数平均值及变异幅度非常接近。同一小孢子再生植株群体内的不同倍性植株的叶绿体数平均值差异极显著。不同倍性植株间的气孔保卫细胞叶绿体数均呈正态分布。本研究提出了一种根据甘蓝类蔬菜气孔保卫细胞叶绿体数进行染色体倍性鉴定的方法,即气孔保卫细胞叶绿体数≤10的为单倍体,10<叶绿体数≤15的为二倍体,>15的为多倍体。该方法经花期形态学观察和根尖染色体计数法验证,其吻合率达93.93%,且此倍性鉴定方法稳定,不受植株生长环境等外界因素影响。【结论】甘蓝类蔬菜游离小孢子再生植株的染色体倍性,可于幼苗期根据气孔叶绿体数目的多少进行鉴定,而且此倍性鉴定方法简单、快捷、经济而又可靠。 展开更多
关键词 甘蓝 青花菜 芥蓝 气孔保卫细胞 叶绿体数 倍性鉴定
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结球甘蓝和青花菜小孢子胚植株再生 被引量:35
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作者 袁素霞 刘玉梅 +4 位作者 方智远 杨丽梅 庄木 张扬勇 孙培田 《植物学报》 CAS CSCD 北大核心 2010年第2期226-232,共7页
结球甘蓝(Brassica oleracea var.capitata)和青花菜(Brassica oleracea var.italica)小孢子胚再生植株频率低是目前影响游离小孢子培养技术有效应用的关键问题之一,研究其小孢子胚植株再生频率的影响因素,提高胚再生植株频率,对促进游... 结球甘蓝(Brassica oleracea var.capitata)和青花菜(Brassica oleracea var.italica)小孢子胚再生植株频率低是目前影响游离小孢子培养技术有效应用的关键问题之一,研究其小孢子胚植株再生频率的影响因素,提高胚再生植株频率,对促进游离小孢子培养技术在甘蓝类蔬菜育种中更好地应用具有重要意义。该文以结球甘蓝中甘11和青花菜TI-111等基因型为试材,对影响游离小孢子胚再生成植株的固体培养基类型、琼脂浓度、胚的类型及胚在液体培养基中的滞留时间等因素进行了研究。结果表明:游离小孢子培养25天的子叶胚在琼脂浓度为1%–1.25%的B5培养基上植株再生频率最高。进一步通过8个不同基因型对上述实验结果进行了验证,结果显示,游离小孢子培养25天的子叶胚在1%琼脂浓度的B5培养基上植株再生频率达77.8%–97.2%。 展开更多
关键词 结球甘蓝 青花菜 子叶胚 游离小孢子培养 植株再生
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芥蓝×甘蓝的F_2群体抽薹期性状QTLs的RAPD标记 被引量:11
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作者 陈书霞 王晓武 +2 位作者 方智远 程智慧 孙培田 《园艺学报》 CAS CSCD 北大核心 2003年第4期421-426,共6页
利用芥蓝C10 0 12 ×甘蓝秋50 Y7杂交组合的F2 作图群体的RAPD标记连锁图进行了 19个农艺学性状的QTL定位研究 ,在 9个连锁群上共定位了 2 8个QTL位点。其中控制抽薹期、开花期、株高 ( 4月 2 7日调查 )的QTL各 3个 ;控制伸展度... 利用芥蓝C10 0 12 ×甘蓝秋50 Y7杂交组合的F2 作图群体的RAPD标记连锁图进行了 19个农艺学性状的QTL定位研究 ,在 9个连锁群上共定位了 2 8个QTL位点。其中控制抽薹期、开花期、株高 ( 4月 2 7日调查 )的QTL各 3个 ;控制伸展度、株高 ( 4月 2 0日调查 )、叶柄长QTL各 2个 ;控制茎高、叶数、叶宽( 4月 2 7日调查 )的QTL各 1个 ;控制叶宽 ( 5月 11日调查 )的QTL有 5个。 展开更多
关键词 芥蓝 甘蓝 F2群体 抽薹期性状 QTLS RAPD标记 连锁图 农艺学性状 开花期 株高
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甘蓝和青花菜杂种小孢子培养 被引量:22
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作者 陈文辉 方淑桂 +2 位作者 曾小玲 朱朝辉 廖晓珍 《热带亚热带植物学报》 CAS CSCD 北大核心 2006年第4期321-326,共6页
对甘蓝(Brassicaoleraceavar.capitata)×青花菜(Brassicaoleraceavar.italica)的20个杂种及相应的父母本进行游离小孢子培养,并对影响甘青杂种小孢子胚胎发生的主要因子进行探讨,适于小孢子培养的培养基为1/2NLN,附加0.5mgL-1NAA、... 对甘蓝(Brassicaoleraceavar.capitata)×青花菜(Brassicaoleraceavar.italica)的20个杂种及相应的父母本进行游离小孢子培养,并对影响甘青杂种小孢子胚胎发生的主要因子进行探讨,适于小孢子培养的培养基为1/2NLN,附加0.5mgL-1NAA、0.05mgL-1BA、5mgL-1AgNO3、0.2mgL-12,4-D和0.1mgL-1活性炭。结果有14个杂种能产生胚状体,诱导率70%;不同杂种间小孢子胚胎发生频率存在很大差异,最高的是绿洲808×夏宝,平均每蕾16.2个胚。诱导杂种胚状体发生的最佳时期是小孢子单核靠边期至双核期,34℃热激2d有利于小孢子细胞对称分裂。在含糖170gL-1的液体培养基中培养3d,添加低糖(含糖110gL-1)的培养液,可显著提高出胚率。 展开更多
关键词 甘蓝 青花菜 杂种 小孢子培养
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