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1.
Protoplasts were isolated from leaves of Lotus corniculatus and Trifolium repens and from cotyledons of L. corniculatus. These protoplasts divided and produced colonies. The plating efficiency of protoplasts of both species was improved when agarose was used as a supporting medium. Plants were regenerated more regularly and in larger numbers from colonies of L. carniculatus than T. repens. The use of a culture line of T. repens that had been selected for its response in culture markedly increased the proportion of protoplast-derived cultures which regenerated shoots. One regenerant of T. repens (P6) was analysed for morphological and cytalogical variation. This plant was abnormal and highly aneuploid with a wide range of chromosome numbers. 相似文献
2.
Protoplasts were isolated from young inflorescence-derived suspension cultures of a japonica rice cultivar ‘Taipei 177’. The isolated protoplasts which were cultured either in liquid, agar on Sea plaque agarose underwent sustained division. Maximum plating efficiency of 1.06% occurred in a medium containing macroelements of KM, microelements and vitamins of B5, 0.5 % Sea plaque agarose, 1.0 mg/l of 2,4-D, and glucose as an osmotic stabilizer. Green and albino plants were regenerated from the protocalli in MS semisolid medium containing 4 mg/l BAP, 0.5 mg/l NAA and 500 mg/l casein hydrolysate (MS18–2). 相似文献
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苎麻悬浮细胞原生质体培养再生植株 总被引:5,自引:0,他引:5
苎麻品种浏阳大叶绿的子叶在含有2,4-D0.5mg/L、KT0.5mg/L的MSB固体培养基上,形成愈伤组织。愈伤组织经3 ̄4次继代培养后作液体振荡培养,产生悬浮细胞系。从悬浮系分离的原生质体,只有以海藻酸钠包埋方式培养在KM8P培养基中,50天左右才能形成肉眼可见的小愈伤组织。该愈伤组织在附加2,4-D0.2mg/L、6-BA0.1mg/L的MSB生长培养基上增殖,然后转入附加6-BA2.0mg 相似文献
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陆地棉原生质体培养与植株再生技术研究 总被引:4,自引:2,他引:2
分离培养陆地棉品种ZDM-3(Gossypium hirsutum L cv ZDM-3)胚性细胞悬浮系的原生质体,通过体细胞胚胎发生成功获得再生植株。试验结果表明,植物生长调节剂组合和碳源对原生质体培养具有重要的影响,添加2,4-D + KT + 1.5%(W/V)葡萄糖+ 1.5%(W/V)麦芽糖的KM8P培养基对于陆地棉原生质体培养的效果较好。激素组合0.46 μmol·L-1 KT + 0.45 μmol·L-1 2,4-D诱导的愈伤组织较松软,分化潜力高;胚性愈伤组织的增殖使用0.93 μmol·L-1 KT + 2.46 μmol·L-1 IBA的激素组合;1.2 μmol·L-1 IBA + 1.39 μmol·L-1 KT有利于体细胞胚胎发生,而MSB-5 + 0.67 μmol·L-1 KT+2.69 μmol·L-1 NAA的培养基适合体细胞胚胎萌发和植株再生。此外,愈伤组织诱导宜用葡萄糖作为碳源,而胚性愈伤组织增殖、保存和胚状体的萌发过程宜使用麦芽糖作碳源。 相似文献
5.
油菜薹茎段髓部原生质体培养再生植株 总被引:2,自引:0,他引:2
以油菜品种“蜀杂6号”的薹茎段髓部为材料分离原生质体,产量一般在每g鲜重2×10~6个左右。在每L附加1.5mgBA、0.4mgNAA、1.5mg2,4-D、200mg水解酪蛋白、250mg木糖、0.3M蔗糖和0.1M葡萄糖的Nitsch培养基中,对原生质体进行液体浅层静置培养。2~7天内原生质体出现第一次分裂,分裂频率最高可达49.2%。培养15天后逐渐形成肉眼可见的小愈伤组织。21天时将培养基改换成每L附加2mgBA、0.2mg 2,4-D和3%蔗糖的MS扩增培养基,45天后可形成5mm左右的愈伤组织。大于2mm的愈伤组织可在每L含有1.0mgBA/3.0mgZT和0.1mgNAA的MS琼脂培养基上分化出芽,分化频率可达36.7%。在1/2MS无激素培养基中,1cm左右的分化芽有85.9%生根,形成完整的植株。 相似文献
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花生不定芽分化及植株再生的研究 总被引:2,自引:2,他引:0
为进一步探索和完善高频植株再生体系并扩大基因型的研究,以8个花生品种为材料,对幼叶不同部位(叶片顶端、中间切段、叶片基部)的切段、子叶、上胚轴和下胚轴外植体,接种到添加1 mg/L NAA和6 mg/L BAP的MSB5培养基上,10天后将形成的愈伤组织转移至添加10 mg/L BAP或添加3 mg/L BAP和1 mg/L ABA的再分化培养基上进行培养,诱导不定芽分化。结果表明,6天叶龄叶片中间切段不定芽分化频率达到了91.4%,上胚轴26.7%,下胚轴12.5%和子叶0%;花生品种8823幼叶外植体获得了的91.4%不定芽分化率,生根植株经驯化后移至砂土中,正常开花结果。因此,幼叶外植体分化不定芽频率明显高于子叶、上胚轴和下胚轴;6天叶龄的叶片中间切段为最佳;不同基因型不定芽分化率存在明显差异。 相似文献
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The genetic determination of the plant regeneration ability of tissue cultures arising from immature embryos was studied using a ‘Chinese Spring’/‘Cheyenne’ substitution series. Plant regeneration proved to be polygenically determined. In tile current experiment the chromosomes 7B, 7D and ID were found to be effective, although the possibility of other chromosome effects cannot be excluded. 相似文献
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Somatic Embryogenesis and Plant Regeneration from Tritordeum 总被引:2,自引:0,他引:2
Regeneration of plants by somatic embryogenesis from immature embryos of hexaploid tritordeum (AABBHchHch, amphiploid Hordeum chilense×Triticum turgidum conv. durum) and durum wheat (Triticum tergidum) was induced on MS medium supplemented with different 2.4-D concentrations. Well-defined embryoids were formed with a high frequency on the scutellar callus from 1 or 2 weeks onwards and plantlets were developed from them. In the best cases from one single explant more than 100 plants could be obtained. Plants were also regenerated by somatic embryogenesis from inflorescences of Hordeum chilense×Triticum turgiditm conv. durum hybrid and its respective hexa-amphiploid. With regard to callus induction and regenerative ability, evident differences between hexa- and octoploid (H. chilense×T. aestivum) tritordeum were found, the latter showing a very low response. 相似文献
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甘蓝型油菜原生质体培养获得再生植株的研究 总被引:7,自引:0,他引:7
以甘蓝型油菜下胚轴游离获得原生质体。经液体-固体双层培养,原生质体分裂、增殖。约四周左右形成细胞团和肉眼可见的小愈伤组织。然后转入固体培养基中增殖。待愈伤组织长到直径约5mm 大小转入分化培养基中。两周后形成绿芽,并诱导生根,获得再生植株。 相似文献
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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
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Genotypic and exogenous factors affecting shoot regeneration from anther callus of linseed (Linum usitatissimum L.) 总被引:1,自引:0,他引:1
Summary The objective of this study was to investigate factors affecting the regeneration capacity of linseed anther culture. Four different environmental conditions in a phytotron were tested with regard to their effects on anther donor plants of cv. Hella. Anther response and shoot regeneration from anther callus was maximal when donor plants were grown in a 16 hrs-day at 14°C day/8°C night temperature. Anthers of four linseed genotypes were cultured on different media. Maximum shoot regeneration was achieved when the induced calli were transferred onto a modified N6 medium containing zeatin (1 mg l-1). Most of the calli regenerated shoots in the second subculture on regeneration media. Shoots were rooted on modified B5 or MS media containing NAA (0.1 mg l-1). Cytological examinations of incubated anthers and root tips of regenerated plants indicated that the anther calli were derived from microspores.Abbreviations B5
Gamborg's (1975) medium
- BAP
6-benzylaminopurine
- 2,4D
dichlorophenoxyacetic acid
- N6
Chu's (1978) medium
- NAA
-naphthaleneacetic acid
- MS
Murashige & Skoog's (1962) medium
- ZEA
zeatin 相似文献
14.
大蕉未成熟雄花接种到胚性愈伤组织诱导培养基中,4~5个月后可诱导出胚性愈伤组织,并可在继代培养基上长期增殖。这些继代培养了6年多的松软易碎的胚性愈伤组织转移到含有0.2 mg/L 6-BA的分化培养基中,可诱导出芽,得到了53株再生植株,这些再生植株可进一步扩大繁殖。组织学切片证明长期继代培养的愈伤组织维持了胚性的状态。 相似文献
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Plant regeneration from anther culture in Canadian cultivars of flax (Linum usitatissimum L.) 总被引:1,自引:0,他引:1
The effect of culture of anthers at 35 °C for one to four days prior to culture at 25 °C in darkness, genotype, anther orientation
on callus induction and shoot regeneration in anther culture of flax was investigated. The influence of type and concentrations
of cytokinins in the regeneration medium on shoot regeneration was also investigated. The results suggested that culture of
anthers at 35 °C prior to continuous culture at 25 °C in darkness did not significantly improve the percentage of anthers
producing calli. However, culture of anthers at 35 °C for one day significantly increased the overall efficiency of regeneration
compared to no culture temperature treatment. Genotypic effects were significant for the percentage of anthers producing calli
and the overall efficiency of regeneration. Anther orientation showed no significant differences. The regeneration medium
containing 4.5 μM zeatin had significantly higher percentage of calli forming shoots than the same basal medium containing
0.01 μM TDZ. The importance of these findings for flax breeding was discussed.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
18.
High frequency of plant regeneration from anther culture in flax, Linum usitatissimum L. 总被引:3,自引:0,他引:3
The effects of induction medium compositions on flax anther culture were investigated in order to improve the efficiency of callus induction and plant regeneration. Anthers were inoculated onto the modified MS medium supplemented with five different combinations of plant growth regulators. The medium containing the combination of 2mg/l 2,4- dichlorophenoxy-acetic acid (2,4-D) and 1 mg/1 6-benzylaminopurine (BAP) produced a significantly higher number of calli forming shoots/100 responded anthers and a significant increase in overall efficiency of regeneration than the same basal medium containing 1 mg/1 a-naphthalene-acetic acid (NAA) and 2 mg/1 BAP (CK). Among the five levels of thiamin hydrochloride tested, the modified MS medium containing 10 mg/1 thiamin hydrochloride significantly increased the number of calli forming shoots/100 responded anthers and the overall efficiency of regeneration compared with the same basal medium containing 0.1 mg/1 thiamin hydrochloride. Maltose concentration had a significant effect on the percentage of anthers producing call, the number of calli forming shoots/100 responded anthers and the overall efficiency of regeneration. The medium containing 6% or 9% maltose produced the highest overall efficiency of regeneration among the five levels of maltose evaluated. Sucrose concentration significantly affected the number of calli forming shoots/100 responded anthers and the overall efficiency of regeneration, and dramatically affected the frequency of microspore-derived plants and the frequency of spontaneous chromosome doubling in microspore-derived plants. The efficiency of doubled haploid line production obtained in this study appears adequate for applied breeding programmes. 相似文献
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降香黄檀愈伤组织培养与植株再生研究 总被引:1,自引:1,他引:1
为了实现降香黄檀工厂化快速繁殖和扩大栽培,并为降香黄檀抗寒基因导入打下一定的基础,以降香黄檀无菌实生苗茎段、叶片、根尖为外植体,MS为基本培养基,对各器官愈伤组织诱导与分化的最适培养基成分进行了研究。结果表明,可从降香黄檀无菌实生苗茎段、叶片、根尖成功地进行愈伤组织培养和植株再生。茎段、叶片、根尖诱导愈伤组织的最适培养基分别为1/2MS+6-BA 1.5 mg/L+NAA 0.10 mg/L、1/2MS+6-BA 0.5 mg/L+NAA 0.10 mg/L和1/4MS+6-BA 1.5 mg/L+NAA 0.05 mg/L;茎段、叶片、根尖的愈伤组织诱导丛生芽最适培养基分别为1/2MS+6-BA 1.5 mg/L+2,4-D 4.0 mg/L、1/2MS+6-BA 1.5 mg/L+2,4-D 3.5 mg/L和1/2MS+6-BA 1.5 mg/L+2,4-D 3.5 mg/L;茎段、叶片、根尖来源的单芽,其最佳生根培养基分别为MS+NAA 1.5 mg/L、MS+NAA 1.0 mg/L和MS+NAA 2.0 mg/L。比较茎段、叶片与根尖的愈伤组织诱导率、丛生芽诱导率和单芽生根率,得出以无菌实生苗根尖作为外植体是降香黄檀愈伤组织培养和植株再生的最佳选择。 相似文献