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1.
We have assayed different combinations of nutrient media and growth regulators to induce callus and plant regeneration from explants of root, shoot and leaf, complete seed, and isolated mature embryo of barley (Hordeum vulgare L. cv. Hassan). The best results were obtained with mature embryo in J25-8 medium supplemented with 2.0 mg dm–3 2,4-dichlorophenoxyacetic acid where about 75 % developed friable calli. Some 80 – 85 % of these calli regenerated barley plants in the same J25-8 medium supplemented with 1.0 mg dm–3 indole-3-butyric acid and 0.1 mg dm–3 kinetin. 相似文献
2.
大麦成熟胚愈伤组织的诱导和植株再生的研究 总被引:2,自引:1,他引:1
以10个大麦优良品种为实验材料,成熟胚为外植体,研究基因型、种子的不同切割方式、培养基、激素等对大麦成熟胚愈伤组织的诱导及植株再生的影响.结果表明,种子纵切后接种出愈率显著高于横切;改良MS培养基能提高出愈率;在愈伤组织诱导过程中,不同品种对激素2,4-D与Dicamba的反应表现不同;初代愈伤组织经过3次继代培养后会转变为两种类型的胚性愈伤组织;不同品种的植株再生在不同浓度有机添加物的分化培养基上表现不同;长时间的继代培养,一些品种在植株再生过程中出现一定数量的白化苗.供试材料均能进行愈伤组织诱导,但是只有部分品种能再生植株.本实验筛选出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87-3175、87-0053、97-4010、97-6004及208813-509. 相似文献
3.
甜高粱再生体系的建立 总被引:2,自引:1,他引:1
利用甜高粱成熟种子和成熟胚作为外植体,通过愈伤组织再生途径建立了植株再生体系。结果表明,成熟种子在添加1.38 g·L^-1脯氨酸、500 mg·L^-1水解酪蛋白和3.0 mg·L^-1 2,4-D的MS培养基上可以较快地诱导出生长状态良好的愈伤组织;成熟胚在添加1.38 g·L^-1脯氨酸、500mg·L^-1水解酪蛋白、2.5 mg·L^-1 2,4-D和0.1 mg·L^-1KT的MS培养基上也能诱导出生长良好的愈伤组织。将愈伤组织转移到MS+1 mg.L^-1 IAA+0.5 mg·L^-1 6-BA培养基上诱导芽,然后再转移到MS+3 mg·L^-1IBA培养基上生根后,可发育成为完整的植株。 相似文献
4.
利用甜高粱成熟种子和成熟胚作为外植体, 通过愈伤组织再生途径建立了植株再生体系。结果表明, 成熟种子在添加1.38 g.L-1脯氨酸、500 mg.L-1水解酪蛋白和3.0 mg.L-1 2, 4-D的MS培养基上可以较快地诱导出生长状态良好的愈伤组织;成熟胚在添加1.38 g.L-1脯氨酸、500 mg.L-1水解酪蛋白、2.5 mg.L-1 2, 4-D和0.1 mg.L-1KT的MS培养基上也能诱导出生长良好的愈伤组织。将愈伤组织转移到MS+1 mg.L-1 IAA + 0.5 mg.L-1 6-BA培养基上诱导芽, 然后再转移到MS+3 mg.L- 1IBA培养基上生根后, 可发育成为完整的植株。 相似文献
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A reliable protocol for plant regeneration from mature embryo derived calli of nine barley (Hordeum vulgare) cultivars has been developed. The auxins 2,4-dichlorophenoxyacetic acid, picloram and dicamba proved effective in inducing
callus from mature embryos of most of the barley cultivars. The induced primary callus was loose, friable and translucent.
It ultimately yielded creamy white and compact callus after 2 - 3 transfers on fresh medium of the same composition. Callus
induction and regeneration capacity were highly cultivar dependent. Addition of a high concentration of picloram (4 mg dm-3) promoted regeneration in 3 cultivars (Tallon, Grimmett and Sloop). In cv. Arapiles, abscisic acid and betaine were crucial
in generating morphogenic callus from the mature embryos. Plants regenerated from these calli were hardy and developed roots
readily when transferred to hormone free medium.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
7.
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
8.
利用成熟种子作为外植体,分析了2,4-D对匍匐翦股颖胚性愈伤组织诱导与植株再生体系的影响,并对体细胞胚的发生过程进行了观察.实验结果表明,在2.0 mg/L 2,4-D 0.1 mg/L 6-BA时胚性愈伤组织的诱导频率最高.随着2,4-D浓度的增加,愈伤组织的诱导和分化能力明显下降.在再生过程中采用1.0 mg/L的6-BA达到了比较好的效果,愈伤组织的再生频率大部分在90%以上.同时发现适当提高肌醇浓度可以使苗长得较为粗壮.在实验中发现匍匐翦股颖的体细胞胚发生过程为球形胚、心形胚、鱼雷胚和子叶胚. 相似文献
9.
本文给出了6个籼型杂交稻亲本成熟胚愈伤组织再生体系建立优化措施。采用6个有重要育种价值的杂交籼稻亲本成熟胚盾片诱导愈伤组织作为分化再生的外植体,通过调节2,4-D浓度、培养基成分、接种方式、激素组合和愈伤组织分化途径,建立适合籼稻遗传转化的再生体系。结果表明,MB培养基是较为合适的愈伤组织诱导培养基类型,6个品种在MB培养基上的愈伤组织诱导率均在60%-80%之间。半粒米的接种方式能够明显提高愈伤组织诱导率,提高幅度达到28.2%。通过调节2,4-D浓度和激素组合,愈伤组织诱导率最高可达到97.9%,两步分化法和适当干燥处理能够提高愈伤组织的分化效率,6品种愈伤组织分化率均在50%-90%之间。初步建立了6个杂交籼稻亲本品种成熟胚愈伤组织的再生体系,为以后遗传转化奠定基础。 相似文献
10.
小麦成熟胚愈伤组织诱导及分化研究 总被引:3,自引:0,他引:3
以2个小麦品种成熟胚为外植体进行离体培养,研究了不同预处理、不同2,4-D浓度及与KT组合、不同蔗糖浓度等因素对愈伤组织诱导及分化的影响。结果表明:4℃低温预处理可提高愈伤组织的出愈率及再生苗率,2个材料的出愈率及再生苗率均达到90%和30%以上;在不同预处理条件下,2,4-D浓度对出愈率及再生苗率的影响与基因型有关,2,4-D浓度为1~2 mg/L更有利于愈伤组织诱导及分化;附加KT能缓解高浓度2,4-D对再生苗率的抑制作用,而对于在1、2 mg/L 2,4-D的培养基中附加KT则不表现这种作用;蔗糖浓度则在30 g/L条件下更有利于愈伤组织诱导。因此通过4℃低温预处理,在MS基本培养基中附加1~2mg/L 2,4-D及30 g/L蔗糖亦可促进小麦成熟胚愈伤组织的诱导和分化。 相似文献
11.
以杂交狼尾草初级愈伤为材料,运用方差分析的方法研究不同因素对杂交狼尾草胚性愈伤诱导率与植株再生率的影响。结果发现,2,4-D和TDZ对杂交狼尾草胚性愈伤诱导影响显著,最佳的胚性愈伤诱导培养基为MS+3.0 mg·L-12,4-D+0.4 mg·L-16-BA+0.2 mg·L-1 TDZ,诱导率为54%;愈伤分化培养基以附加0.1 mg·L-1 TDZ+0.2 mg·L-16-BA+3.2 mg·L-1 CuSO4的 MS培养基为最佳,再生率为68.33%,褐化率为8.33%;分化培养基中添加0.8~3.2mg·L-1的CuSO4均能促进杂交狼尾草愈伤组织分化,而添加AgNO3对杂交狼尾草愈伤分化无显著影响。该技术为离体诱变获得杂交狼尾草低温种质材料奠定了基础。 相似文献
12.
Somatic Embryogenesis and Plantlet Regeneration from Callus of Mature Zygotic Embryos of Masson Pine
Embryogenic cultures were initiated from mature zygotic embryos of masson pine (Pinus massoniana Lamb. ) on DCR medium supplemented with 2, 4-D 10 mg/L, KT and BA each at 4 mg/L. Pale and translucent calli with early stage proembryos were maintained and multiplicated on DCR medium supplemented with 2, 4-D 1.0 mg/L KT and BA each at 0.4 mg/L. Robust late-stage proembryos were obtained when the calli were cultured on DCR medium containing 9000 mg/L myo-inositol. Abscisic acid and activated charcoal promoted the formation of cotyledonary embryos at the highest frequency of 35.1 %. Mature somatic embryos could germinate and develop further into plantlets when they were isolated and cultured on a hormone-free DCR medium. 相似文献
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以可在黑龙江地区露地越冬的5个现代月季(Rosa chinensis)品种为实验材料,分别以其无菌苗的叶片和茎段为外植体,研究了愈伤组织诱导及植株再生方法。实验结果表明:5个寒地月季品种的叶片和茎段均可诱导出愈伤组织,2,4-D诱导愈伤组织的效果较好,高浓度的细胞分裂素不适合用于月季叶片和茎段愈伤组织的诱导;TDZ在月季愈伤组织分化培养过程中具有重要作用,光照培养可促进月季愈伤组织的分化,愈伤组织的分化能力随着继代次数的增加呈下降趋势。该实验成功地从2004-8和2004-9(2个月季品种)愈伤组织中诱导出再生植株,其愈伤组织的分化率分别为45%和38%。 相似文献
15.
Callus was induced from mature embryos of Alstroemeria cv. Butterflycultured on MS medium supplemented with 2·0 or 4·0mg dm3 2,4D or picloram and incubated at 25°Cin the dark. The effect of auxin concentration and precultureof embryos was studied. Callus was capable of regeneration aftertransfer to MS medium containing 4.0 mg dm°3 BAP. Shootsand whole plantlets were regenerated. The effect of growth regulators,used in the callus induction medium and the regeneration medium,on plant regeneration was studied Key words: Alstroemeria, callus, plant regeneration. 相似文献
16.
阳桃胚乳愈伤组织诱导和不定芽发生的研究 总被引:5,自引:0,他引:5
首次成功建立阳桃胚乳组织培养并获得胚乳再生植株。胚乳愈伤组织诱导以培养基MS 2,4-D2.0mgL^-1 BA0.2mgL^-1的效果最好,诱导频率可达94.7%,愈伤组织乳白色,结构致密,生长旺盛;将其接种在培养基MS ZT3.0mgL^-1 NAA0.2mgL^-1上,愈伤组织由乳白色致密型转变为淡绿色致密型,进而形成绿色芽点,分化出不定芽,分化频率可达73.3%;胚乳植株在培养基MS ZT2.0-2.5mgL^-1 NAA0.05mgL^-1上进行壮苗和营养繁殖。 相似文献
17.
野牛草幼穗愈伤组织的诱导及植株再生 总被引:5,自引:0,他引:5
以野牛草[Buchloe dactyloides(Nutt.)Engelm.]幼穗为外植体,建立了愈伤组织诱导、继代培养和植株再生体系。结果表明,雌穗比雄穗难以脱分化形成愈伤组织;小于8mm雄幼穗在2mg/L2,4-D培养基上的愈伤组织诱导率为80.0%~86.8%;添加10mg/L AgNO3对愈伤组织诱导率影响不明显,但可改善愈伤组织质量。2mg/L 2,4-D结合0.1mg/L 6-BA的培养基有利于愈伤组织的继代培养;继代超过3次、继代间隔超过3周,愈伤组织分化能力明显下降。雄穗愈伤组织在含1.0mg/L 6-BA培养基上,弱光条件下分化出芽的频率较高,达31.8%~35.0%;附加3%麦芽糖既可减轻褐化程度,又利于丛生芽的分化。分化苗在1/2MS 0.3mg/L IBA培养基上的生根率为62.5%。 相似文献
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植物名称桂竹香(Cheiranthus cheiri)。2 材料类别下胚轴,子叶。3 培养条件桂竹香种子用自来水洗净后,以70%酒精进行表面消毒30 s,再以0.1%升汞溶液消毒12 min,然后用无菌水洗4次。将种子接种在MS基本培养基上,1周后切下子叶和下胚轴,分别接种在MS+2,4-D 1mg·L-1(单位下同)+NAA 1+6-BA 1和MS+2,4-D 3+6-BA0.2~0.5上诱导形成愈伤组织。分化和生长培养基为MS+6-BA3.0+NAA0.2;生根培养基为MS+IBA 0.5。蔗糖浓度为3%,琼脂浓度为0.7%,培养温度为(25±2)℃。每天光照10 h,光照度1 500~2 000 lx。4 生长与分化情况子叶和下胚轴接种到诱导培养基上,20 d左右开始形成愈伤组织。愈伤组织接种到分化培养基以后,经20 d左右的培养,开始出现许多绿点,继而出现绿芽。将单个绿芽切下接种在生长培养基上,2 周后形成丛生芽。 将芽切下接种在生根培养基上,25 d以后开始生根,生根率100%。当根的数量达到2~4条,长度在0.5 cm以上时即可移栽。移栽前先打开瓶塞,在室温下炼苗2~3 d,取出小苗,洗去根部培养基,移植于土中,每天用小喷雾器喷水一次,成活率可达90%以上。5 意义与进展桂竹香是十字花科桂竹香属草本花卉。据我们观察,它的抗油菜菌核病性较好,其组织培养尚未见报道。本文中桂竹香子叶和下胚轴的愈伤组织诱导和植株再生技术的建立,对通过体细胞杂交将桂竹香的抗菌核病性导入油菜,可能有一定的参考价值,同时也为桂竹香的保纯提供了一条值得考虑的途径。 相似文献
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黄花补血草愈伤组织的诱导和植株再生 总被引:8,自引:3,他引:5
以黄花补血草(Limonium aureum(L.)Hill.)无菌苗为材料,研究了不同激素配比条件下不同外植体愈伤组织的诱导及植株再生.结果表明,黄花补血草无菌苗叶片、叶柄和幼根均可作为离体培养的外植体,但叶片和叶柄的诱导率明显高于幼根.将外植体接种于分别添加0.5 mg/L NAA或1.0 mg/L 2,4-D或0.3-0.5 mg/L 6-BA 0.5-2.0 mg/L NAA的MS培养基上,经过14-28 d培养后,可脱分化产生乳白色、红色或浅绿色颗粒状或致密愈伤组织,频率达到70%以上.在MS 0.3 mg/L 6-BA 1.0 mg/L NAA培养基上,红色和绿色颗粒状愈伤组织经过1-2次继代后,均可分化产生不定芽,进而形成丛生芽,分化率达到100%.将高约3 cm的丛生芽切下,接种于分别添加0.5 mg/L IAA或0.5 mg/L IBA的1/2 MS培养基上可产生不定根,获得完整的再生植株. 相似文献