甘蓝型油菜组培苗生根培养体系的优化
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摘要
为解决油菜组织培养中组培苗生根细短、移栽不易成活的问题,以甘蓝型半冬性油菜转化p CAMBIA1301空载体的再生植株为材料,以生根时间、根长、根重、苗高、苗重、总重、生根数、生根率等指标,评价以MS和B5为基础培养基,分别使用琼脂糖、琼脂粉及植物凝胶为凝固剂时再生苗的生根效率,得到了两种基础培养基的最适凝固剂组合;在以上两种组合的培养基中分别加入不同浓度蔗糖,比较生根情况和成本,认为含20g/L蔗糖的B5琼脂粉培养基为最优配方。利用优化后的生根培养基配方,可以极大地促进油菜组培苗的生根长度和数目,提高后期移栽成活率。
In rapeseed tissue culture,tenuous and short roots lead to low survival rate of transplanted. To modify the rooting system and to ensure related study,shoots regenerated from a semi-winter oilseed rape cultivar transformed with empty p CAMBIA1301 vector were used. Rooting time,root length,shoot height,shoot weight and plantlet biomass were investigated to assess rooting efficiency of basal medium MS and B5 combined with agar,agarose and phytagel as gelling agent. Results showed that,considering the cost,B5 agar medium containing 20 g/L sucrose were the optimal formula. The optimized rooting medium formula could increase root length and number of in vitro plantlets,and thus enhance the survival rate after transplanting.
In rapeseed tissue culture,tenuous and short roots lead to low survival rate of transplanted. To modify the rooting system and to ensure related study,shoots regenerated from a semi-winter oilseed rape cultivar transformed with empty p CAMBIA1301 vector were used. Rooting time,root length,shoot height,shoot weight and plantlet biomass were investigated to assess rooting efficiency of basal medium MS and B5 combined with agar,agarose and phytagel as gelling agent. Results showed that,considering the cost,B5 agar medium containing 20 g/L sucrose were the optimal formula. The optimized rooting medium formula could increase root length and number of in vitro plantlets,and thus enhance the survival rate after transplanting.
引文
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[6]Sun F,Fan G,Hu Q,et al.The high-quality genome of Brassica napus cultivar‘ZS11’reveals the introgression history in semi-winter morphotype[J].Plant J,2017,92(3):452-468.
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[14]Lin N,Liu L,Yin J,et al.Establishment of transformation system using inbred line of yellow-seeded Brassica napus[J].Acta Agronomica Sinica,2013,38(12):2 185-2 191.
[15]何俊平,涂小云.不同培养基配方对铁皮石斛生根培养的影响[J].江苏农业科学,2013,41(2):57-59.
[16]曹剑,黄志伟.不同培养基配方对白及组培苗生根培养的影响[J].四川林业科技,2016,37(3):96-99+53.
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[19]王炳涵,丰景,杨万年,等.农杆菌介导的甘蓝型油菜“中双11号”的遗传转化[J].安徽农业科学,2012,40(24):11 957-11 960.
[20]廖志强,邬贤梦,孙娟,等.甘蓝型油菜下胚轴一步不定芽再生培养及遗传转化应用[J].分子植物育种,2015,13(4):793-799.
[21]Liu F,Xiong X,Wu L,et al.Bra LTP1,a lipid transfer protein gene involved in epicuticular wax deposition,cell proliferation and flower development in Brassica napus[J].PLo S One,2014,9(10):e110272.
[22]徐银,宋刚,曹正,等.不同配方凝固剂对马铃薯脱毒试管苗生长及其微型薯生产的影响[J].中国马铃薯,2015,29(6):325-328.
[23]吴清平,孙永,蔡芷荷,等.微生物固体培养基凝固剂研究进展[J].微生物学通报,2006(5):145-149.
[24]张巧,汪静儿,林君,等.不同凝固剂对陆地棉体细胞胚胎发生和植株再生的影响[J].棉花学报,2010,22(1):3-9.
[25]李龙.龙须菜琼脂糖的提取技术研究[D].福州:福建农林大学,2013.
[26]韩继成,陈霜莹,章德明.Phytagel R作为固化剂获得苹果叶片高频率植株再生[J].河北农业科学,2006(4):36-39.
[27]张翠平,李琳琳,韦悦,等.3种常用培养基对颠茄毛状根生长与次生代谢产物积累的影响[J].西南大学学报(自然科学版),2017,39(6):36-41.
[28]车志.厚朴组织培养技术研究[D].福州:福建农林大学,2009.
[29]严华兵,杨丽涛,李俊玲,等.不同蔗糖浓度对山薯组培苗形态发生途径的影响[J].热带作物学报,2011,32(7):1 325-1 329.
[30]胡江琴,王利琳,庞基良,等.绿豆下胚轴正反插条件下无机盐和蔗糖对下胚轴不定根发生的影响[J].植物生理学通讯,2002,38(3):228-230.
[31]张硕,黄豫谦,张飞雄.蔗糖对小麦根生长和形态学的影响[J].安徽农业科学,2012,40(35):16 987-16 988.
[32]De Faria T,Rodrigues N,Oliveira V.R,et al.In vitro dendrobium nobile plant growth and rooting in different sucrose concentrations[J].Horticultura Brasileira,Brasilia,2004,22(4):780-783.
[2]Ooms G,Bains A,Burrell M,et al.Genetic manipulation in cultivars of oilseed rape(Brassica napus)using Agrobacterium[J].Theor Appl Genet,1985,71(2):325
[3]De Block M,De Brouwer D,Tenning P.Transformation of Brassica napus and Brassica oleracea using Agrobacterium tumefaciens and the expression of the bar and neo genes in the transgenic plants[J].Plant Physiol.1989,91(2):694-701.
[4]Braatz J,Harloff H J,Mascher M,et al.CRISPR-Cas9targeted mutagenesis leads to simultaneous modification of different homoeologous gene copies in polyploid oilseed rape(Brassica napus)[J].Plant Physiol,2017,174(2):935-942.
[5]Chalhoub B,Denoeud F,Liu S,et al.Plant genetics.Early allopolyploid evolution in the post-Neolithic Brassica napus oilseed genome[J].Science,2014,345(6199):950-953.
[6]Sun F,Fan G,Hu Q,et al.The high-quality genome of Brassica napus cultivar‘ZS11’reveals the introgression history in semi-winter morphotype[J].Plant J,2017,92(3):452-468.
[7]Rani T,Yadav R C,Yadav N R,et al.Genetic transformation in oilseed brassicas-A review[J].Indian J Agric Sci,2013,83(4):367-373.
[8]Schr9der M,Dixelius C,Rahlen L,et al.Transformation of Brassica napus by using the aad A gene as selectable marker and inheritance studies of the marker genes[J].Physiologia Plantarum,1994,92(1):37-46.
[9]Cardoza V,Stewart C N.Increased Agrobacterium-mediated transformation and rooting efficiencies in canola(Brassica napus L.)from hypocotyl segment explants[J].Plant Cell Reports,2003,21(6):599-604.
[10]Halfhill M D,Richards H A,Mabon S A,et al.Expression of GFP and Bt transgenes in Brassica napus and hybridization with Brassica rapa[J].Theor Appl Genet,2001,103(5):659-667.
[11]Khan M R,Rashid H,Quraishi A.High frequency shoot regeneration and Agrobacterium mediated DNA transfer in Canola(Brassica napus L.)[J].Plant Cell Tissue Organ Cult,2003,75:223-231.
[12]Maheshwari P,Selvaraj G,Kovalchuk I.Optimization of Brassica napus(canola)explant regeneration for genetic transformation[J].New Biotechnol,2011,29(1):144-55.
[13]Liu X X,Lang S R,Su L Q,et al.Improved Agrobacterium-mediated transformation and high efficiency of root formation from hypocotyl meristem of spring Brassica napus'Precocity'cultivar[J].Genet Mol Res,2015,14(4):16 840-16 855.
[14]Lin N,Liu L,Yin J,et al.Establishment of transformation system using inbred line of yellow-seeded Brassica napus[J].Acta Agronomica Sinica,2013,38(12):2 185-2 191.
[15]何俊平,涂小云.不同培养基配方对铁皮石斛生根培养的影响[J].江苏农业科学,2013,41(2):57-59.
[16]曹剑,黄志伟.不同培养基配方对白及组培苗生根培养的影响[J].四川林业科技,2016,37(3):96-99+53.
[17]陈春福,赖钟雄.二乔杜鹃组培苗生根培养体系的优化研究[J].园艺与种苗,2012(12):36-39.
[18]潘刚,周永明.甘蓝型油菜遗传转化的研究进展[J].中国油料作物学报,2003,25(3):92-100.
[19]王炳涵,丰景,杨万年,等.农杆菌介导的甘蓝型油菜“中双11号”的遗传转化[J].安徽农业科学,2012,40(24):11 957-11 960.
[20]廖志强,邬贤梦,孙娟,等.甘蓝型油菜下胚轴一步不定芽再生培养及遗传转化应用[J].分子植物育种,2015,13(4):793-799.
[21]Liu F,Xiong X,Wu L,et al.Bra LTP1,a lipid transfer protein gene involved in epicuticular wax deposition,cell proliferation and flower development in Brassica napus[J].PLo S One,2014,9(10):e110272.
[22]徐银,宋刚,曹正,等.不同配方凝固剂对马铃薯脱毒试管苗生长及其微型薯生产的影响[J].中国马铃薯,2015,29(6):325-328.
[23]吴清平,孙永,蔡芷荷,等.微生物固体培养基凝固剂研究进展[J].微生物学通报,2006(5):145-149.
[24]张巧,汪静儿,林君,等.不同凝固剂对陆地棉体细胞胚胎发生和植株再生的影响[J].棉花学报,2010,22(1):3-9.
[25]李龙.龙须菜琼脂糖的提取技术研究[D].福州:福建农林大学,2013.
[26]韩继成,陈霜莹,章德明.Phytagel R作为固化剂获得苹果叶片高频率植株再生[J].河北农业科学,2006(4):36-39.
[27]张翠平,李琳琳,韦悦,等.3种常用培养基对颠茄毛状根生长与次生代谢产物积累的影响[J].西南大学学报(自然科学版),2017,39(6):36-41.
[28]车志.厚朴组织培养技术研究[D].福州:福建农林大学,2009.
[29]严华兵,杨丽涛,李俊玲,等.不同蔗糖浓度对山薯组培苗形态发生途径的影响[J].热带作物学报,2011,32(7):1 325-1 329.
[30]胡江琴,王利琳,庞基良,等.绿豆下胚轴正反插条件下无机盐和蔗糖对下胚轴不定根发生的影响[J].植物生理学通讯,2002,38(3):228-230.
[31]张硕,黄豫谦,张飞雄.蔗糖对小麦根生长和形态学的影响[J].安徽农业科学,2012,40(35):16 987-16 988.
[32]De Faria T,Rodrigues N,Oliveira V.R,et al.In vitro dendrobium nobile plant growth and rooting in different sucrose concentrations[J].Horticultura Brasileira,Brasilia,2004,22(4):780-783.