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1.
植物组织培养再生相关基因鉴定、克隆和应用研究进展 总被引：2，自引：0，他引：2
离体植物组织体细胞胚胎发生是一个复杂的无性繁殖过程，依次经历外源植物激素信号应答、已分化细胞的脱分化、静止细胞的再分裂以及特定组织、器官原基或分生组织的形成等，是多个基因在外界因素刺激下协调、有序表达和互作的结果，不但受培养基中植物激素和营养成分的影响，也与外植体的生理状态关系密切。本文综述了外源激素和内源激素在植物组织培养中的作用，以及外源激素对内源激素的调节功能；重点介绍了5类与植物体细胞胚胎发生有关的候选基因，包括体细胞胚胎发生相关类受体蛋白激酶、阿拉伯葡聚糖酶、亚硝酸还原酶、生长素结合蛋白和抗氧化酶。再生相关基因的利用不但有助于提高植物组织培养植株再生率和遗传转化率，而且有助于获得安全型转基因植物，在基因工程育种中具有潜在应用前景。不同植物和同种植物不同外植体组织培养中调控体细胞胚胎发生的主效基因可能不同，关键再生相关基因的克隆和功能鉴定是今后需要加强的方向。 相似文献
2.
3.
低酚陆地棉直接体细胞胚胎发生和植株再生 总被引：2，自引：0，他引：2
选用低酚陆地棉无菌苗下胚轴为材料进行全固体组织培养,直接诱导获得了胚性愈伤组织,并进一步分化为再生植株.结果表明,激素是影响棉花直接体细胞胚胎发生的重要因素.MSB培养基中添加2,4-D有利于愈伤组织的形成,却不能直接诱导获得胚性愈伤组织.MSB培养基中添加IBA和BA也不能直接诱导获得胚性愈伤组织.MSB培养基附加适当浓度的IBA和KT能直接诱导出胚性愈伤组织.最适激素组合(1.0 mg/L IBA,0.5 mg/L KT)能使诱导棉花下胚轴产生大量胚性愈伤组织,并且在3个月内就可肉眼观察到不同发育时期的胚.MSB培养基中附加1.0 g/L谷氨酰胺和0.5 g/L天门冬酰胺有利于胚萌发成苗.本研究建立了简便高效的棉花直接体细胞胚胎发生和植株再生培养体系,从胚性愈伤组织诱导到植株再生约需5～6个月时间. 相似文献
4.
【目的】研究脱落酸(Abscisic acid, ABA)对棉花体细胞胚胎发生过程中下胚轴脱分化和再分化的影响,优化体细胞胚胎发生体系和初步解析脱落酸调控棉花体细胞胚胎发生分子机制。【方法】以棉花品种中棉所24(CCRI 24)下胚轴为外植体,设置5个ABA浓度0、0.02、0.04、0.06、0.08μmol·L^-1,分别以A0、A1、A2、A3、A4表示,添加至MSB(MS培养基+B5维生素)培养基诱导愈伤和胚性愈伤,研究ABA对棉花下胚轴初始细胞脱分化、愈伤组织诱导和胚性愈伤组织诱导的影响。【结果】ABA促进下胚轴初始细胞脱分化;显著提高愈伤组织的脱分化率和增殖率;0.02μmol·L^-1ABA显著提高胚性愈伤分化率,0.04~0.08μmol·L^-1ABA显著降低胚性愈伤分化率。ABA处理后胚性愈伤和非胚性愈伤的增殖率均显著提高且质地受到影响。0.02~0.08μmol ABA处理下,LBD和LBD在愈伤起始期上调表达。0.02μmol·L^-1ABA处理下,在愈伤增殖早期和中期BBM、LEC1和AGL15上调表达,愈伤增殖后期FUS3、LEA、ABI3基因上调表达。【结论】脱落酸调控的棉花体细胞胚胎发生与相关标记基因的时空性表达密切相关,这些基因表达水平的增加是ABA调控愈伤和胚性愈伤分化的分子基础。 相似文献
5.
《棉花学报》2021,(1)
【目的】研究脱落酸(Abscisic acid, ABA)对棉花体细胞胚胎发生过程中下胚轴脱分化和再分化的影响,优化体细胞胚胎发生体系和初步解析脱落酸调控棉花体细胞胚胎发生分子机制。【方法】以棉花品种中棉所24(CCRI 24)下胚轴为外植体,设置5个ABA浓度0、0.02、0.04、0.06、0.08μmol·L~(-1),分别以A0、A1、A2、A3、A4表示,添加至MSB(MS培养基+B5维生素)培养基诱导愈伤和胚性愈伤,研究ABA对棉花下胚轴初始细胞脱分化、愈伤组织诱导和胚性愈伤组织诱导的影响。【结果】ABA促进下胚轴初始细胞脱分化;显著提高愈伤组织的脱分化率和增殖率;0.02μmol·L~(-1)ABA显著提高胚性愈伤分化率,0.04～0.08μmol·L~(-1)ABA显著降低胚性愈伤分化率。ABA处理后胚性愈伤和非胚性愈伤的增殖率均显著提高且质地受到影响。0.02～0.08μmol ABA处理下,LBD和LBD在愈伤起始期上调表达。0.02μmol·L~(-1)ABA处理下,在愈伤增殖早期和中期BBM、LEC1和AGL15上调表达,愈伤增殖后期FUS3、LEA、ABI3基因上调表达。【结论】脱落酸调控的棉花体细胞胚胎发生与相关标记基因的时空性表达密切相关,这些基因表达水平的增加是ABA调控愈伤和胚性愈伤分化的分子基础。 相似文献
6.
茉莉酸在调控棉花体细胞胚增殖方面有重要作用,而丙二烯氧化物合酶(AOS)是植物茉莉酸合成途径中的关键酶。作者鉴定出在胚发育过程中表达的AOS基因,这些基因编码区长度均在1500 bp左右,被定位在5条染色体上,有2个基因含有内含子。对其启动子元件分析发现,它们可能受激素和胁迫的调控在体细胞胚胎发生中起作用。荧光定量分析发现,这些基因在棉花不同组织中的表达模式主要分为3类,GhAOS1和GhAOS6在棉花胚发育过程中表达量相对较高,GhAOS2和GhAOS3在棉花愈伤和胚性愈伤中表达量相对较高,GhAOS4和GhAOS5在茎和叶中表达水平相对较高,暗示这些基因的表达有时空特异性,且GhAOS1和GhAOS6可能在棉花胚发育中发挥重要作用。亚细胞定位分析显示,大部分被定位在过氧化物酶体中,可能参与调控光呼吸、体细胞胚成熟和萌发。上述分析为进一步研究AOS基因家族在棉花体细胞胚发育中的功能奠定了基础。 相似文献
7.
《分子植物育种》2021,19(7):2265-2272
DNA甲基化是表观遗传修饰的途径之一,在维持生物基因组稳定性与调控基因转录和表达中起着重要作用。为明确棉花愈伤组织分化过程中基因组DNA的甲基化变异模式以及甲基化图谱,本研究利用全基因组亚硫酸盐测序(WGBS)技术对棉花愈伤组织分化(非胚性与胚性)过程中的DNA甲基化模式进行了全面比较。全基因组DNA甲基化分析表明,在棉花愈伤组织分化过程中非胚性愈伤组织的mCG与m CHG所占比率高于胚性愈伤组织,而不对称m CHH的比率则低于胚性愈伤组织;非胚性愈伤组织的mCG、m CHG、mCHH及mC的平均甲基化水平均低于胚性愈伤组织。基因组甲基化区域分析表明,棉花愈伤组织分化过程中基因间、启动子和基因下游(转录终止位点下游2 kb)的甲基化水平较高,同时胚性愈伤组织的基因间、启动子及基因下游的平均甲基化水平均高于非胚性愈伤组织。因此,本研究分析了棉花愈伤组织全基因组DNA甲基化水平,为进一步研究棉花体细胞胚胎发生的表观遗传机制提供了依据。 相似文献
8.
棉花组织培养直接胚胎发生和植株再生 总被引：11，自引：1，他引：11
选用陆地棉(GossypiumhirsutumL.)品种中棉所12,首次直接诱导获得棉花体细胞胚胎发生,并获得了再生植株。结果表明,激素是影响棉花体细胞胚胎直接发生的重要因素,单独使用ZT可直接诱导获得棉花胚性愈伤组织和胚状体,2,4-D的添加虽有利于愈伤组织的形成,但却没能直接诱导获得胚性愈伤组织,IAA的添加削弱了ZT的诱导效果。下胚轴、子叶和胚根直接诱导获得胚胎发生的能力不同,其中以子叶的分化能力最强,胚根次之,下胚轴较差。实验中,棉花体细胞胚胎直接发生的最高频率为11.42%,占诱导获得总愈伤组织的44.44%。 相似文献
9.
外源激素对棉花体细胞胚胎发生及发育的调控作用 总被引：16，自引：6，他引：10
选用11种激素研究了外源激素对棉花胚性愈伤组织增殖、胚胎发生和发育的调控作用。结果表明，不同激素对棉花胚性愈伤组织增殖、胚胎发生与发育的影响不同。除2，4-D和BA对棉花胚性愈伤组织的增殖影响不大外，其它激素对棉花胚性愈伤组织的增殖均具有抑制作用，且具有一定的时间效应，同时还受基因型的影响。激素对棉花体细胞胚的形成和发育的影响极大，2，4-D既抑制了体细胞胚的形成，又抑制了体细胞胚的发育；TDZ的作用与2，4-D相似，显著抑制了体细胞胚的形成，且诱导获得的体细胞胚均停留在球形胚阶段；GA也抑制了体细胞胚的形成，且不利于体细胞胚的成熟与萌发；4BU-30对棉花体细胞胚形成与发育的影响不大。其它7类生长素类物质和细胞分裂素类物质对棉花体细胞胚的形成均具有促进作用，且依IBA、ABA、IAA、BA、KT、ZT、2iP次序增强，其总胚数为对照的1．193～3．852倍；其中2iP的促进作用最大，可使产生的体细胞胚数提高2．852倍。 相似文献
10.
ＴＤＺ是一种高效低毒棉花脱叶剂，它在棉花组织培养中表现出独特的生理作用。ＴＤＺ的添加促进了外植体的伸长和保绿，有利于外植体脱分化和胚性愈伤组织的诱导。但是ＴＤＺ不利于愈伤组织的保持和体细胞胚的发育，必须把在含ＴＤＺ培养基上诱导产生的愈伤组织转移到不含任何激素的培养基上，才能发育成成熟胚并再生出植株。ＴＤＺ的使用能诱导根的分化。不同外植体、不同基因型以及不同ＴＤＺ浓度之间在棉花组织培养中的表现均有差异。 相似文献
11.
二倍体、四倍体小麦成熟胚组织培养研究初报 总被引：2，自引：0，他引：2
以栽培二粒小麦(Triticum dicoccum Schuble)、硬粒小麦(Triticum durum Desf.)以及普通六倍体小麦祖先种野生二粒小麦(Triticum dicoccoides Kom.)、野生一粒小麦(Triticum aegilopides Bal.)4个基因型麦属植物成熟胚(MEs)为外植体,对成熟胚愈伤组织诱导、分化及植株再生的组织培养效果做了初步研究,筛选出适合于非六倍体小麦成熟胚组织培养的基因型.结果表明,不同基因型成熟胚愈伤组织的诱导、分化及植株再生差异显著,具有很强的基因型效应和基因型依赖性.其中栽培二粒小麦的成熟胚表现出较好的组织培养特性,它的愈伤组织诱导率、分化率、成苗率以及组培效率分别为95.00%、90.00%、32.40%、27.70%,显著高于其它基因型.二倍体、四倍体小麦成熟胚组织培养特性的研究以及组织培养基因型的筛选,为遗传转化体系的建立、小麦功能基因的分离和克隆,以及小麦分子育种工作奠定了基础. 相似文献
12.
长江和黄河流域棉区棉花品种体细胞胚胎发生和植株再生比较研究 总被引：6，自引：0，他引：6
选用我国长江和黄河流域棉区各10个棉花品种以及珂字201和YZ1共22个基因型,研究和优化棉花体细胞胚胎发生的相关条件参数。在此基础上,比较了两生态区棉花品种的体细胞再生能力。结果表明, 在IBA+KT的激素组合下,多数基因型有较强再生能力;在愈伤组织诱导时期,铵态氮是必需的,而在愈伤组织继代分化时期,一定浓度的硝态氮则能促进分化;没有铁盐不能诱导出愈伤组织,而铁盐浓度为56 mg L-1时有利于胚分化;长江流域和黄河流域棉花品种的植株再生潜力没有明显差别,但长江流域的品种体细胞胚胎发生所需的时间长。本文首次获得了鄂抗棉3号、5号、鄂棉20、鄂棉23、豫棉9号、豫早73、豫棉12、豫棉1221等8个品种的体细胞胚胎发生和植株再生。 相似文献
13.
The cotyledon and hypocotyl of eight varieties of alfalfa( Medicago sativa L. )are used to study the effect of different cultivation procedures and genotypes on embryogenic callus and embryoid induction,plant development and transplant of alfalfa.The varieties of alfalfa with high embryoid induction rate and high differentiation rate are screened. A high frequency regeneration system for alfalfa tissue culture is established:which is benifical to the gene transformation of alfalfa. 相似文献
14.
Somatic embryogenesis (SE) is a critical step leading to plant regeneration in tissue culture of many plant species. The objective
of the present study was to analyze the inheritance of SE in cotton (Gossypium hirsutum L.) using leaf petioles as explants. A high embryogenic callus (HEC)—producing line, W10, was selected by petiole callus
culture from a commercial Chinese cotton cultivar CRI24 and crossed with a non embryogenic line, TM-1 and a low embryogenic
(LEC) commercial Chinese cotton cultivar, CRI12, respectively. The parental lines, F1 and F2 were grown in field conditions for sources of leaf petioles as explants. The F1 plants were similar to the HEC parent in embryogenic callus (EC) induction, indicating that high EC ability is dominant.
The classical Mendelian analysis showed that the high EC ability in the HEC line W10 is controlled by two independent dominant
genes with complementary effect, designated Ec
1
and Ec
2
, while the LEC line CRI12 contains one dominant gene Ec
2
. A joint segregation analysis confirmed that SE ability in cotton is controlled by two major genes with epistatic effects
along with other polygenes. A SSR marker analysis identified three quantitative trait loci (QTLs) on two linkage groups, one
of which harbored a major QTL (qEc1) which is assigned to the major gene Ec
1
. This qualitative and quantitative genetic study has provided an incentive to fine map the genes responsible for SE towards
the isolation of the SE genes in cotton. 相似文献
15.
玉米成熟胚胚性愈伤组织的诱导、高频再生及转化的研究 总被引：4，自引：0，他引：4
以玉米自交系CML295、CML304和18-599R的成熟胚为外植体, 结合幼胚离体培养方法, 探讨并优化了成熟胚来源的胚性愈伤组织诱导及继代培养方法。对其愈伤组织的形态和组织切片的研究结果显示, 继代过程可产生良好的Ⅱ型胚性愈伤组织。在分化培养中分别获得68.6%、75.4%和84.8%的高频再生率, 每愈伤组织块成苗数分别为2.45、2.43和2.75。利用基因枪法转化pCAMBIA1301质粒后的GUS瞬时表达效率分别为57.9%、62.5%和73.1%, 转化pCAMBIA1303质粒后检测GFP的瞬时表达效率分别为23.3%、40%和45.5%。以上3种基因型成熟胚来源的愈伤组织转化率与其对应的幼胚来源的胚性愈伤组织转化率相似。这一技术体系为玉米的遗传改良和功能基因组研究提供了重要的技术平台。 相似文献
16.
D. Sihachakr R. Haïcour J.M. Cavalcante Alves I. Umboh D. Nzoghé A. Servaes G. Ducreux 《Euphytica》1997,96(1):143-152
The application of new techniques for improvement of sweet potato crops, particularly including the exploitation of somaclonal
variation, gene transfer by genetic transformation and somatic hybridization, requires the control of plant regeneration from
tissue cultures. Shoots can easily be regenerated from explants of stems, petioles, leaves and roots, while callus cultures
do not produce any shoots. The potential of somatic embryogenesis and plant regeneration via embryogenesis was evaluated for
10 cultivars of sweet potato. Protocols for plant regeneration from cultured protoplasts have also been developed. Since mesophyll
was resistant to enzyme digestion, fragments of stems and petioles, callus and cell suspensions were used as source of protoplasts
of sweet potato. Series of transfers of protoplast-derived calluses, particularly those which had been obtained from in vitro
plants, to media containing a high level of zeatin resulted in successful formation of shoots in only two sweet potato cultivars.
In addition, the embryogenic potential was irreversibly lost through protoplast culture, since protoplasts isolated from embryogenic
cell suspensions developed into non-embryogenic callus. Consequently, an alternative protocol is being successfully developed
to improve plant regeneration from cultured protoplasts of sweet potato, involving first root formation from which shoots
can then be regenerated. Preliminary evaluation in field conditions in Gabon revealed that plants regenerated from cultured
protoplasts exhibited a great genetic variability in their growth and tuber formation in particular.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
17.
燕麦愈伤组织诱导和分化再生影响因素的研究 总被引：9，自引：0，他引：9
以燕麦3个品种(系)幼胚、幼穗为材料,通过组织培养方法,研究了基因型、培养基、外植体对愈伤组织诱导、继代和分化再生的影响。结果表明,对愈伤组织的诱导,基因型和培养基起重要作用,对幼胚作用极显著,对幼穗作用显著;外植体不同也影响愈伤组织形成,随培养基成分改变而变化,且幼穗较幼胚更易培养;2,4-D浓度影响愈伤组织生长和胚性愈伤组织形成,3 mg/L 2,4-D有利于愈伤生长,促进胚性愈伤形成;草莜一号幼穗愈伤组织有很强的继代能力,继代培养330 d仍具有46.58%的分化率,该材料在组织培养和基因工程研究中具有很大潜力。 相似文献
18.
C.E. van Schaik C. van der Toorn M.J. De Jeu C.J.J.M. Raemakers R.G.F. Visser 《Euphytica》2000,115(1):17-26
The successful application of plant biotechnology to Alstroemeria improvement will largely depend on the availability of an efficient regeneration/transformation system. Regeneration in Alstroemeria is accomplished from nodular embryogenic callus initiated from zygotic embryos. Histological studies of embryogenic callus
initiation from 4-weeks old cultured ovules revealed that the outermost layers of the protoderm of the embryogenic nodules
divided to form either a new nodule or aproembryo. Transient gene expression after particle bombardment of nodular embryogenic
callus was optimized using DNA of pAHC25. The highest β-glucuronidase expression was found when the GUS gene was under control of the maize ubiquitin promoter, the target tissue was placed 5 cm below the microcarrier launch assembly
and when the rupture disc-breakage point was between 650–900 psi. Kanamycin blocked regeneration of somatic embryos, however,
did not block growth of nodular embryogenic callus. With phosphinothricin both callus growth and regeneration were blocked.
Bombardment of nodular embryogenic callus with DNA of pAHC25 combined with selection on medium containing phosphinothricin
resulted in putative transgenic chimeric. Friable calli were selected from nodular embryogenic callus and used to initiate
suspensions. These cell suspensions were subjected to transformation by particle bombardment using DNA of pAHC25 and resulted
in a stable transformed friable callus line after selection based on luciferase activity. Even after 2 years of maintenance
this callus line was luciferase positive and the Polymerase Chain Reaction analysis demonstrated the presence of the introduced
gene in this friable callus line.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
19.
The behaviour of diverse Triticum genotypes in the tissue culture response of mature embryo callus was compared, and factors affecting tissue culture response were studied in this paper. Significant differences were detected in callus induction, embryogenic callus differentiation, plantlet regeneration and culture efficiency when mature embryos of 31 plants of different Triticum species were compared. These were the main wheat cultivars of the Chinese northern winter‐type wheat region and breeding lines (Triticum aestivum L.), durum wheat (Triticum durum Desf.), cultivable emmer wheat (Triticumdicoccum Schuble) and the common wheat progenitors Triticum dicoccoides and Triticum aegilopides. The genotype dependency was particularly high in tissue culture of mature embryos of these Triticum genotypes. The efficiency of induction, differentiation and regeneration of mature embryos callus was high in genotypes selected out. Mature embryo‐derived callus of HB341, TS021, SN2618, T. dicoccum, HB188, and T9817 showed better tissue culture response than the other genotypes. Plantlets can be regenerated from mature embryo‐derived callus of 31 genotypes, saving on growth facility resources and time required for the collection of other explants, and providing a solid basis for the genetic transformation and molecular plant breeding of Triticum plants. 相似文献
20.
Production of embryogenic tissues and regeneration of transgenic plants in cassava (Manihot esculenta Crantz) 总被引：3，自引：0，他引：3
Nigel J. Taylor Munyaradzi V. Masona Rosa Carcamo Thao Ho Christian Schöpke Claude M. Fauquet 《Euphytica》2001,120(1):25-34
Disorganised embryogenic tissues have been utilised as target tissues for transgene insertion and transgenic plant regeneration
in cassava (Manihot esculenta). The production of friable embryogenic callus in fourteen geographically diverse cassava cultivars, from which eleven were
established as embryogenic suspension cultures, is reported. Embryogenic tissues were similar in nature in all cultivars tested
although there was variation in the time required to generate friable callus and the growth rates of suspension cultures.
Regeneration of plants has been achieved from eight cultivars but varied significantly in efficiency, with cv. TMS 60444and
Line 2 from Zimbabwe being the most responsive. Tissues from the remaining eight cultivars became arrested at globular and
torpedo stages of regeneration indicating that they most likely process an inherent ability to produce plants but require
further research to allow this to be realised. Significant numbers of transgenic plants containing transgenes for putative
resistance to important viral diseases of cassava in addition to visual marker genes have been regenerated. Transgenic plants
from three the cultivars TMS 60444, Bonoua Rouge and M.Col 1505 were recovered after particle bombardment of embryogenic suspension
cultures. Correlation's have been made between abnormal leaf morphology and plant vigour with the use of embryogenic suspension
cultures for transgene insertion. As an result friable embryogenic callus is now being successfully utilsed as the target
tissue for genetic transformation and plant regeneration at ILTAB.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献