共查询到19条相似文献,搜索用时 187 毫秒
1.
玉米优良自交系成熟胚再生体系的建立 总被引:4,自引:0,他引:4
选用生产上广泛应用的10个玉米优良自交系,用幼胚通过组织培养研究其再生特性,结果表明:玉米自交系基因型间的培养能力有较大的差异,自交系178的再生率高达78%。在此基础上以其中的178玉米优良自交系为材料,研究了影响玉米成熟胚再生的各种因素,结果表明:高浓度的2,4-二氯苯氧乙酸(2,4-D)(4.0 mg/L)是诱导愈伤组织必须的;在继代培养基中添加适量的2,4-D(2.0 mg/L)、6-苄基嘌呤(6-BA)(0.2 mg/L)和硝酸银(10 mg/L)显著增加胚性愈伤组织的形成;在分化培养基中添加0.5 mg/L 6-BA有利于提高愈伤组织的分化频率。该再生体系的建立,为以成熟胚为受体系统的遗传转化体系的建立奠定了基础。 相似文献
2.
不同杂种优势类群玉米幼胚愈伤组织的诱导及植株再生特性的研究 总被引:19,自引:1,他引:18
以Reid、唐四平头和其它种质等3个杂种优势类群共19份玉米自交系为试验材料,以玉米幼胚作为外植体,研究了基因型、培养基和激素对玉米幼胚愈伤组织的诱导及植株再生的影响,结果表明供试材料均能进行愈伤组织的诱导,但是仅有12个自交系能再生植株。N6和改良N6培养基有助于提高愈伤组织的质量及其生长速度,2,4-D在愈伤组织的诱导中起着关键性作用。在诱导培养基中添加0.2mg/L的6-BA或KT会使胚性愈伤组织的诱导频率下降以及降低愈伤组织的质量。在胚状体诱导培养基中添加1mg/L的KT能促进绿苗的分化,但是浓度过高会使丛生苗分化过多。此外,通过对不同杂种优势类群自交系玉米幼胚培养特性的分析,发现在唐四平头类群的4个自交系中,黄早四的绿苗分化率仅为0.5%,其它3个自交系不能再生植株。但是,从Reid和其它种质类群的供试自交系中筛选出了胚性愈伤组织的诱导频率和绿苗分化率均较高的、适合于遗传转化的受体材料,如3189/4380、4380/陕综5、8103、先早17、18-599红、18-599白、501、178和冀53。 相似文献
3.
不同杂种优势类群玉米幼胚愈伤组织的诱导及植株再生特性的评价 总被引:5,自引:1,他引:4
以Reid、唐四平头和其他种质等3个杂种优势类群共30份玉米自交系为实验材料,以玉米幼胚作为外植体,研究基因型、培养基、激素、继代培养次数对玉米幼胚愈伤组织诱导及植株再生的影响.研究结果表明供试材料均能进行愈伤组织诱导,但是只有部分自交系能再生植株.通过对不同杂种优势类群自交系玉米幼胚培养特性的分析,从Reid、唐四平头和其他种质类群的供试自交系中,筛选出了胚性愈伤组织诱导频率和绿苗分化率均较高、适合于遗传转化的受体材料,如黄野四/京24//C108/黄野四选系、黄早四/先早17//吉853选系、R43//黄野四/711选系、京7、京7黄、3189/4380选系、4380/陕综5选系、8103、先早17、18-599红、501、178和冀53. 相似文献
4.
甘蓝型油菜子叶和下胚轴再生植株无性系建立 总被引:4,自引:1,他引:3
以甘蓝型油菜(Brassica napus L.)豫油2号和6257的子叶和下胚轴为材料,在不同激素配比的MS培养基上诱导出了愈伤组织。将经过继代的部分愈伤转入分化培养基,分化结果表明:除基因型、外植体和分化培养基的激素配比对分化率有影响外,诱导愈伤培养基的激素配比对分化率也至关重要。豫油2号的子叶和下胚轴在最适诱导培养基(ZT 1+NAA0.5+2,4-D 0.2 mg/L)和最适分化培养基(ZT4+IAA 0.2 mg/L)组合中的愈伤分化率分别为12.5%和75%;6257的子叶和下胚轴在其最适诱导培养基(KT 2+NAA1+2,4-D 0.2 mg/L)和最适分化培养基(6-BA 4+IAA 0.02 mg/L)组合中的愈伤分化率分别为50%和37.5%。将其最适诱导培养基中的愈伤组织继代达8个月以上,建立了不同继代愈伤的再生植株无性系。 相似文献
5.
探索华山松(Pinus armandii)体细胞胚胎发生技术对其实施规模化无性繁殖和开展遗传转化具有重要意义。本文以1/2LM为基本培养基, 通过激素调节等措施对华山松的胚性愈伤组织诱导和幼胚的离体培养技术进行了初步研究。研究结果: 胚性愈伤组织诱导率最高可达52.71%, 但愈伤组织继代培养后没有体细胞胚胎的分化; 首次从其子叶期的幼胚中直接诱导出具有根和茎的完整植株, 诱导率达92%以上。文章确认了采集的幼胚发育状态对胚性愈伤组织的诱导有重要影响, 并对诱导的培养条件等进行了探讨。 相似文献
6.
华山松胚性愈伤组织诱导与幼胚离体培养 总被引:5,自引:0,他引:5
探索华山松(Pinus armandii)体细胞胚胎发生技术对其实施规模化无性繁殖和开展遗传转化具有重要意义。本文以1/2LM为基本培养基,通过激素调节等措施对华山松的胚性愈伤组织诱导和幼胚的离体培养技术进行了初步研究。研究结果:胚性愈伤组织诱导率最高可达52.71%,但愈伤组织继代培养后没有体细胞胚胎的分化;首次从其子叶期的幼胚中直接诱导出具有根和茎的完整植株,诱导率达92%以上。文章确认了采集的幼胚发育状态对胚性愈伤组织的诱导有重要影响,并对诱导的培养条件等进行了探讨。 相似文献
7.
《中国野生植物资源》2017,(4)
以玉米幼胚为外植体,研究了烯效唑对玉米幼胚愈伤组织诱导以及愈伤组织分化成植株的影响。结果表明,烯效唑(S-3307)的浓度越高玉米幼胚胚芽的生长受抑制作用越强,胚性愈伤组织发生率越高,有的基因型幼胚对过高浓度的烯效唑表现出不适反应,以至于胚性愈伤转化成非胚性愈伤;不同基因型的玉米幼胚在不同浓度的烯效唑影响下愈伤组织诱导率差异较大;烯效唑的浓度越高胚性愈伤组织分化率也高,但不同基因型对烯效唑的反应不尽相同。 相似文献
8.
9.
具有高再生性且对农杆菌敏感玉米自交系的筛选 总被引:2,自引:0,他引:2
以70个高代玉米自交系的幼胚为材料在NB培养基上进行离体培养,通过多次继代选择,从中筛选出了胚性愈伤组织诱导率高、克隆能力强的10个自交系,将这10个自交系的一部分胚性愈伤组织用作绿苗再分化,研究其再生能力;另一部分用农杆菌C58和GV3301转化,利用植物载体中携带的gus和g彦报告基因的瞬时表达为指标研究玉米基因型对农杆菌的敏感性。绿苗再分化结果表明,10个自交系都具有绿苗再分化能力,其中自交系6010、6038、6015、6051和6060的绿苗再分化力相对较高,分别为61.11%、31.94%、45%、33.33%和156.94%。gus瞬时表达率方差分析结果表明:玉米基因型对农杆菌C58的敏感性存在极显著差异,在自交系6034、6038的胚性愈伤组织上没有检测到gus瞬时表达,即这两个基因型对农杆菌C58不敏感,不能被其转化;其他7个基因型的平均gus瞬时表达率均大于70%,说明这7个基因型对农杆菌C58敏感。GFP荧光检测结果表明,在自交系6034、6038、6069和6010的胚性愈伤组织上没有检测到gyp瞬时表达,在其他6个自交系上检测到绿色荧光蛋白的表达。因此,认为自交系6015、6051和6060是对农杆菌GV3301敏感且具有高再生能力的玉米转基因受体材料;自交系6051、6010、6015、6060和6050是再生能力强且对农杆菌C58敏感的玉米转基因受体材料。 相似文献
10.
11.
In this paper, we describe the transformation of regenerable maize tissues by electroporation. In many maize lines, immature zygotic embryos can give rise to embryogenic callus cultures from which plants can be regenerated. Immature zygotic embryos or embryogenic type I calli were wounded either enzymatically or mechanically and subsequently electroporated with a chimeric gene encoding neomycin phosphotransferase (neo). Transformed embryogenic calli were selected from electroporated tissues on kanamycin-containing media and fertile transgenic maize plants were regenerated. The neo gene was transmitted to the progeny of kanamycin-resistant transformants in a Mendelian fashion. This showed that all transformants were nonchimeric, suggesting that transformation and regeneration are a single-cell event. The maize transformation procedure presented here does not require the establishment of genotype-dependent embryogenic type II callus or cell suspension cultures and facilitates the engineering of new traits into agronomically relevant maize inbred lines. 相似文献
12.
Peng Jiao Ruiqi Ma Zhuo Qi Zhenzhong Jiang Siyan Liu Jing Qu Shuyan Guan Yiyong Ma 《Phyton》2020,89(1):121-130
This research uses the immature embryos of inbred maize lines
(GSH9901, Hi01, Hi02, and Chang 7-2) as receptor materials to establish the callus
induction system. These inbred lines provide the receptor materials for the genetic
regeneration of maize and the verification of the genetic functions of maize. The
factor experiment and orthogonal experiments were used to investigate the impacts
of different genotypes, immature embryo size, shield orientation, 2, 4-D
concentration, proline concentration, and folic acid concentration on the induction
rate of embryogenic callus tissue. A sensitivity experiment testing glyphosate (Bar)
and an antibiotic (Cefotaxime sodium) were also conducted. The results indicate
that the immature embryos of inbred maize line GSH9901 were the most effective
for callus tissue induction, and the immature embryos with a length of 1.6-2.0 mm
produce the best result. The upward shield face is more successful for the
formation of induced callus. Using orthogonal analysis, we found that the optimal
combination for the induction system was A3 (2,4-D concentration 0.25 mg mL-1
),
B1C3 (proline concentration 0.8 mg mL-1
), and D2 (folate Concentration 0.5 mg
mL-1) and the induction rate reached 84%. We found that cold storage at 4 °C for 1
d is more conducive for the formation of embryogenic callus than the other
treatments tested. The sensitivity experiment for callus tissue screening revealed
the critical concentration of glyphosate to be 10 mg ml-1
, and the critical
concentration of antibiotic is 250 mg ml-1
. Using this combination of glyphosate
and antibiotic resulted in regenerated plants. This study established the optimal
conditions for immature embryo callus tissue induction in maize. 相似文献
13.
High-frequency plant regeneration through callus initiation from mature embryos of maize (<Emphasis Type="Italic">Zea Mays</Emphasis> L.) 总被引:10,自引:0,他引:10
An efficient maize regeneration system was developed using mature embryos. Embryos were removed from surface-sterilized mature seeds and sliced into halves. They were used as explants to initiate callus on induction medium supplemented with 4.0 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D). The induction frequency of primary calli was over 90% for all inbred lines tested. The primary calli were then transferred onto subculture medium supplemented with 2.0 mg l–1 2,4-D. Following two biweekly subcultures, embryogenic calli were formed. Inclusion of a low concentration (0.2 mg l–1) of 6-benzylaminopurine (BA) in the subculture medium significantly promoted the formation of embryogenic callus. The addition of silver nitrate (10 mg l–1) also supported an increased frequency of embryogenesis. The embryogenic callus readily formed plantlets on regeneration medium supplemented with 0.5 mg l–1 BA. The regenerated plantlets were transferred to half-strength Murashige and Skoog medium supplemented with 0.6 mg l–1 indole-3-butyric acid to develop healthy roots. The regenerated plantlets were successful on transfer to soil and set seed. Using this system, plantlets were regenerated from seven elite maize inbred lines. The frequency of forming green shoots ranged from 19.8% to 32.4%. This efficient regeneration system provides a solid basis for genetic transformation of maize.Abbreviations BA 6-Benzylaminopurine - 2,4-D 2,4-Dichlorophenoxyacetic acid - IBA Indole-3-butyric acid - KT KinetinCommunicated by M.C. Jordan 相似文献
14.
骨干玉米自交系丹598遗传再生体系的建立 总被引:2,自引:0,他引:2
目的:以玉米骨干自交系丹598的幼胚为外植体,诱导愈伤组织建立遗传再生体系。方法:探讨胚龄、培养基种类、2,4-D浓度对愈伤组织诱导的影响。结果:在授粉后16~18 d,2,4-D浓度为2.0 mg/L时诱导最佳;设置N6、NB、改良NB、MS、MB等5种培养基,筛选出改良NB培养基为最佳诱导培养基;分化培养基中添加1 mg/L激动素、0.5 mg/L 6-卞基嘌呤和0.5 mg/L萘乙酸能促进绿苗分化和根系生长。结论:建立了玉米自交系丹598的优良再生体系,为以后的基因转化工作打下了良好基础。 相似文献
15.
Summary Induction of androgenesis in maize leads to heterogeneous callus types in terms of chromosome number and regeneration ability. In order to obtain homogeneous cell suspensions from androgenetic material we initiated maize cultures from single microspore-derived structures. Cultures were established either by transfer of previously selected type-II callus or by transfer of single aggregates directly into suspension medium. During establishment no selection was performed. Independent suspension lines were analysed with respect to their chromosome number, their embryogenic capacity, and their regeneration ability. Cytological analysis revealed that most of the cultures tested showed a constant chromosome number over a period of 18 months, indicating a homogeneous constitution. Within regenerable lines no difference in embryogenic capacity could be observed when smaller (1mm) and larger (4mm) aggregates were compared. Furthermore, their homogeneous character was confirmed by the regeneration of only green or albino plants from the individual lines. Chromosome analysis of regenerants by root tip cytology showed that their numbers corresponded to that of the suspension they were regenerated from. Similar results showing the homogeneous nature of the lines were obtained in protoplast culture and regeneration experiments.Abbreviations PEG
polyethylenglycol, 2,4-D, 2,4-dichlorophenoxy-acetic acid
- BAP
6-benzylaminopurine 相似文献
16.
Chun-Lai Zhang Dong-Fang Chen Marie Kubalakova Jian Zhang Nigel W. Scott Malcolm C. Elliott Adrian Slater 《Plant Cell, Tissue and Organ Culture》2008,93(2):209-221
Efficient regeneration via somatic embryogenesis (SE) would be a valuable system for the micropropagation and genetic transformation
of sugar beet. This study evaluated the effects of basic culture media (MS and PGo), plant growth regulators, sugars and the
starting plant material on somatic embryogenesis in nine sugar beet breeding lines. Somatic embryos were induced from seedlings
of several genotypes via an intervening callus phase on PGo medium containing N6-benzylaminopurine (BAP). Calli were mainly induced from cotyledons. Maltose was more effective for the induction of somatic
embryogenesis than was sucrose. There were significant differences between genotypes. HB 526 and SDM 3, which produced embryogenic
calli at frequencies of 25–50%, performed better than SDM 2, 8, 9 and 11. The embryogenic calli and embryos produced by this
method were multiplied by repeated subculture. Histological analysis of embryogenic callus cultures indicated that somatic
embryos were derived from single- or a small number of cells. 2,4-dichlorophenoxyacetic acid (2,4-D) was ineffective for the
induction of somatic embryogenesis from seedlings but induced direct somatic embryogenesis from immature zygotic embryos (IEs).
Somatic embryos were mainly initiated from hypocotyls derived from the cultured IEs in line HB 526. Rapid and efficient regeneration
of plants via somatic embryogenesis may provide a system for studying the molecular mechanism of SE and a route for the genetic
transformation of sugar beet. 相似文献
17.
18.
C. L. Armstrong J. Romero-Severson T. K. Hodges 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1992,84(5-6):755-762
Summary The frequency of initiation of friable, embryogenic callus from immature embryos of the elite maize inbred line B73 was increased dramatically by introgression of chromosomal segments from the inbred line A188 through classical backcross breeding. Less than 0.2% of the immature B73 embryos tested (5 of 3,710) formed embryogenic callus. The breeding scheme consisted of six generations of backcrossing to B73 with selection at each generation for high frequency initiation of embryogenic cultures. BC6 individuals were selfed for four generations to select homozygous lines. The average embryogenic culture initiation frequency increased to 46% (256/561). Nearly all (91%) of the embryos from one BC6 S4 plant formed embryogenic cultures. RFLP analysis was used to determine the locations and effects of the introgressed A188 chromosomal segments. Five segments were retained through at least the fifth backcross generation. The hypothesis that one or more of these five regions contains genes controlling somatic embryogenesis in maize was tested using an F2 population of the cross A188 X Mo17. A set of five DNA markers (three of them linked) explained 82% of the observed phenotypic variance for percentage of immature embryos forming embryognic callus. Four of the five markers were located in or near introgressed A188 chromosome segments.The region marked by probe c595 on the long arm of chromosome 9 was highly associated with several measures of in vitro culture response (percent embryogenic embryos, plants per embryo, and plants per embryogenic embryo). We propose that there is a major gene (or genes) in this region in A188 that promotes embryogenic callus initiation and plant regeneration in B73, Mo17, and probably many other recalcitrant inbred lines of maize. 相似文献
19.
Production of transgenic maize from bombarded type II callus: Effect of gold particle size and callus morphology on transformation efficiency 总被引:8,自引:0,他引:8
Bronwyn R. Frame Hongyi Zhang Suzy M. Cocciolone Lyudmila V. Sidorenko Charles R. Dietrich Sue Ellen Pegg Shifu Zhen Patrick S. Schnable Kan Wang 《In vitro cellular & developmental biology. Plant》2000,36(1):21-29
Summary Here we present a routine and efficient protocol for year-round production of fertile transgenic maize plants. Type II callus
derived from maize Hi II immature zygotic embryos was transformed using the PDS 1000/He biolistic gun and selected on bialaphos.
In an effort to improve the transformation protocol, the effects of gold particle size and callus morphology on transformation
efficiency were investigated. Reducing gold particle size from 1.0 μm or 0.6 μm resulted in a significant increase in the
rate of recovery of bialaphos-resistant clones from Type II callus. The average transformation efficiency of pre-embryogenic,
early embryogenic and late embryogenic callus did not vary significantly. Rates of transformation, regeneration and fertility
achieved for Type II callus are summarized and compared to those achieved for greenhouse- and field-derived immature zygotic
embryos. 相似文献