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1.
仙客来体细胞胚发生和发育过程中淀粉粒的动态变化 总被引：2，自引：0，他引：2
以仙客来(Cyclamen persicum Mill.)开花植株的新生叶片为外植体,诱导筛选胚性愈伤组织,并使其进一步发育成体细胞胚.组织切片观察结果表明:胚性愈伤组织由胚性和非胚性细胞组成,胚性细胞多以胚性细胞团的形式存在,胚性细胞团起源于诱导的胚性决定细胞.体细胞胚起源于单个胚性细胞,经多细胞原胚、球形胚、心形胚和鱼雷胚等时期发育成完整植株.在体细胞胚的发生发育过程中,淀粉粒出现4次积累高峰,分别为胚性细胞、球形胚、早期鱼雷胚和成熟胚发育成完整的小植株时期,淀粉代谢与体细胞胚发生、发育及小植株的形态建成密切相关. 相似文献
2.
将香榧(Torreya grandis‘Merrillii’)的未成熟合子胚置于SH+0.1 mg·L-1 NAA+500 mg·L-1AC+3%蔗糖+0.5 g·L-1 Gln培养基上暗培养45 d,诱导产生半透明颗粒状胚性愈伤组织;将胚性愈伤组织转入SH+20 g·L-1 PEG+10 mg·L-1 ABA培养基中暗培养3个月诱导体细胞胚。采用碘—碘化钾染色和石蜡切片技术对体胚起源、形态发育与细胞组织学进行了观察。结果表明:胚性愈伤组织起源于合子胚胚轴表皮或皮层细胞的对称分裂。胚性愈伤组织含有两类细胞,一种是细胞质浓厚、细胞核大、体积小的圆形胚性细胞,另一种是高度液泡化拉长的细胞。胚性愈伤组织包含由这两种细胞构成的原胚团Ⅰ、原胚团Ⅱ和原胚团Ⅲ,以及一些游离细胞。原胚团Ⅲ在无植物生长调节剂的SH基本培养基上形成原胚,原胚接入成熟培养基,历经球形、棒状、心形、鱼雷形胚后发育成子叶胚。将子叶胚转入萌发培养基后胚根伸长,胚芽发育长出针叶,形成完整的再生植株。同时在离体培养中合子胚胚柄处退化的裂生多胚也能重新发育形成胚体。在初生体胚发育过程中表面常伴有次生体细胞胚的形成。 相似文献
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4.
以水芋幼叶为外植体,应用均匀设计法筛选其最适合的胚性愈伤组织及胚性细胞复合体诱导、体细胞胚胎发育及植株再生的培养基,建立了水芋体细胞胚胎发生体系.对不同阶段培养材料的形态结构及超微结构的观察证明了水芋体细胞胚胎的发育过程.结果表明:水芋叶片胚性愈伤组织及胚性细胞复合体诱导最适宜培养基为:LS+6-BA 0.5 mg/L+2,4-D 2.0mg/L,诱导率为98.5%;体胚发育及植株再生最适宜的培养基为:LS+6-BA1.0 mg/L+NAA0.5mg/L+IAA 1.0 mg/L,体胚萌发率为96%,萌发的体胚在发育培养基上继续培养25 d后全部发育成完整植株. 相似文献
5.
以水芋幼叶为外植体,应用均匀设计法筛选其最适合的胚性愈伤组织及胚性细胞复合体诱导、体细胞胚胎发育及植株再生的培养基,建立了水芋体细胞胚胎发生体系。对不同阶段培养材料的形态结构及超微结构的观察证明了水芋体细胞胚胎的发育过程。结果表明：水芋叶片胚性愈伤组织及胚性细胞复合体诱导最适宜培养基为：LS＋6-BA0．5mg／L＋2,4-D2．0mg／L,诱导率为98．5％;体胚发育及植株再生最适宜的培养基为：IS＋6-BA1．0mg／L＋NAA0．5mg／L＋IAA 10mg／L,体胚萌发率为96％,萌发的体胚在发育培养基上继续培养25d后全部发育成完整植株。 相似文献
6.
以‘Red Lion’朱顶红(Hippeastrum vittatum)的鳞片为外植体进行体细胞胚诱导和形态学与组织学的胚性鉴定。结果表明,鳞片在MS+2 mg·L~(-1) BA+1 mg·L~(-1) NAA+2 mg·L~(-1) TDZ+30 g·L~(-1)蔗糖培养基上胚性愈伤组织诱导率可达90.63%,而不添加TDZ的培养基上胚性愈伤组织诱导率为0。胚性愈伤组织可呈白色松散、透明膨松、透明致密、微棕色松脆和翠绿色松脆等类型;非胚性愈伤组织呈微棕色致密状。胚性愈伤组织中可明显观察到球形胚、心形胚和棒状胚3个发育时期。体细胞胚在不添加植物生长调节剂的MS培养基上可发育形成芽和根,长成完整小植株。 相似文献
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以不同生态型的黄瓜（Cucumis sativus L.）为材料，利用不同基因型建立黄瓜体细胞胚再生体系，并在此基础上对
黄瓜体细胞胚再生体系进行了优化。结果表明：华南型黄瓜9101 和D4 能够诱导出正常的胚状体。以筛选出的9101 为材料，
进一步通过不同组织部位、激素配比、暗培养等对体细胞胚再生体系进行优化，认为子叶节是最佳的外植体，其胚性愈伤
组织诱导的最适培养基为MS + 2.00 mg·L-1 2，4-D，诱导率可达100.00%，出胚的最适培养基为MS+0.10 mg·L-1 2，4-D + 0.5
mg·L-1 KT，其出胚率可达33.33%。此外，在愈伤诱导培养前期给予暗培养条件能提高愈伤诱导率、出胚率及正常再生植株
的获得。对体细胞胚的形态学观察发现，黄瓜体细胞胚发源于子叶节表面的胚性愈伤组织，经历球形胚、心形胚、鱼雷形胚
和子叶胚等时期，最终发育成完整的黄瓜植株。 相似文献
8.
以东方百合‘Siberia’的无菌苗叶柄为试材,诱导产生初始分生结节组织。其后对分生结节组织进行分化再生采用体式显微镜观察和石蜡切片分析的方法对诱导得到的类体细胞胚和体细胞胚再生结构进行形态学和组织学观察。百合类体细胞胚结构呈球形至心形结构或芽状,组织内部主要由薄壁细胞组成,与母体组织存在着明显的维管组织联系,具有多个维管组织中心,其外形与体细胞胚极为相似。百合体细胞胚发源于分生结节表面新生的胚性愈伤组织,经历球形、椭圆形和子叶形胚阶段发育形成完整的植株,在整个发育过程中与母体愈伤组织在整个发育过程中无维管组织联系,且它的根芽两极同时发生,相互联系。研究结果表明,两种再生结构虽然具有极相似的外观形态,但是在发育方式和内部组织结构上截然不同。 相似文献
9.
【目的】提高番木瓜体细胞胚发生的同步性及其植株再生率,为番木瓜大量快繁、细胞工程和分子育种技术研发提供技术基础。【方法】以紫晖110～120 d果实的未成熟合子胚为外植体,经胚性愈伤组织诱导、液体悬浮培养和体细胞胚发生过程,建立均质胚性细胞悬浮系和高效植株再生技术体系,重点比较了不同质量浓度2,4-二氯苯氧乙酸(2,4-Dichlorophenoxyacetic acid,2,4-D)对胚性愈伤组织诱导、不同质量浓度6-苄基氨基嘌呤(6-Benzylaminopurine,6-BA)+萘乙酸(1-naphthlcetic acid,NAA)组合和活性炭(activated carbon,AC)对子叶期体细胞胚萌发与生根的影响。【结果】4 mg·L-12,4-D可诱导62.86%未成熟合子胚形成胚性愈伤组织。经5个继代周期的筛选培养,可建立由单细胞和小细胞团组成的均质胚性细胞悬浮系。采用液体培养方式能诱导大量球形胚的形成,被转移至含5 g·L-1AC的半固定培养基成熟培养30 d后,可获得大量子叶期体细胞胚。在含0.4 mg·L-1<... 相似文献
10.
《果树学报》2019,(12)
【目的】明确幼胚愈伤诱导和胚抢救最佳时期,为杧果高效体胚发生体系构建及胚抢救技术有效利用提供依据。【方法】以'热农1号'杧果为材料,探讨幼胚不同发育时期和植物生长调节剂对愈伤组织诱导的影响,同时比较了幼胚不同发育时期对幼胚萌发的影响。【结果】授粉后25 d幼胚处于球形胚期,30 d处于心形胚期,35 d处于鱼雷胚期,40 d之后处于子叶胚期。授粉后40 d早期子叶胚愈伤组织诱导率显著高于其他时期;早期子叶胚在含有3.0 mg·L~(~(-1))2,4-D、1.0 mg·L~(~(-1))KT(或0.5 mg·L~(~(-1))KT及0.5 mg·L~(~(-1))ZT)培养基(PM3和PM4)中愈伤组织诱导率最高,为54.4%～62.2%,显著高于其他培养基。授粉后35 d鱼雷胚的萌发率最高,为47.2%,显著高于其他时期。愈伤组织能经体胚发生途径正常成苗,幼胚萌发后也能正常发育成苗。【结论】'热农1号'杧果幼胚愈伤诱导最适宜的时期是早期子叶胚,胚抢救最适宜的时期是鱼雷胚阶段。 相似文献
11.
扁桃杧(Mangifera persiciformis)体胚发生及再生体系建立 总被引：1，自引：0，他引：1
以扁桃杧(Mangifera persiciformis Wu&Ming)未成熟珠心组织为外植体,建立体胚发生及再生体系,同时对幼苗进行茎尖染色体计数和胚根组织形态学观察。结果表明,在改良的B5基本培养基+2,4-D1.0mg·L-1+Gln400mg·L-1+6%蔗糖上培养4～5周后可诱导胚性愈伤组织分化。继代培养基与成熟培养基交替培养能有效降低胚性愈伤组织的褐化并保持旺盛的分化能力。培养3～4个月后,大部分体胚均能发育成熟,26.03%的体胚畸形。体胚在改良B5培养基+Gln400mg·L-1+4%蔗糖上的萌发率较低,仅为8.39%。次级体胚以直接体胚发生方式于萌发体胚的下胚轴产生。幼苗的生根不理想,生长极为缓慢;其茎尖染色体数目为2n=2x=40;胚根形态学上端内部维管组织解体,愈伤化,结构松散。 相似文献
12.
《Scientia Horticulturae》2005,105(1):117-126
The objectives of the present work were to study the embryogenic competence of floral tissues of Feijoa sellowiana and to investigate the influence of plant growth regulators on somatic embryo induction and development in order to establish a somatic embryogenesis protocol starting from somatic tissues. Petals, stamens and ovaries of floral buds were cultivated onto LPm basal medium supplemented with different levels of 2,4-D, Picloram, 2-iP, Kin and BAP. The highest embryogenic callus induction was obtained with Picloram (10 μM) and Kin (1 μM). Rates of embryogenic calluses induction in stamens and petals were significantly affected by PGRs. Embryogenic calluses were transferred to the same medium, supplemented with gradually reduced levels of PGRs-free medium. After 60 days in suspension cultures with 2,4-D (1 μM) and 2-iP (1 μM) calluses were transferred to PGR-free medium. After 30 days it was observed the development of globular somatic embryos on the surface of 18% of friable calluses previously induced with Picloram (10 μM) and Kin (1 μM). Only embryogenic calluses derived from stamens gave rise to this morphogenetic pattern.Torpedo and cotyledonary somatic embryos transferred to PGR-free culture medium were converted to complete plantlets. This is the first report of somatic embryogenesis in this species starting from somatic tissues. 相似文献
13.
Mahanom Jalil Wong Wei Chee Rofina Yasmin Othman Norzulaani Khalid 《Scientia Horticulturae》2008,117(4):335-340
Somatic embryogenesis is a routine method in obtaining mass-production of plantlets especially in Musa spp. Somatic embryogenesis is a process characterized by a series of morphological changes leading to plant regeneration. Therefore it is important to identify stages in somatic embryogenesis through morphological and histological characteristics. In this study, stages of somatic embryogenesis through morphohistological study could differentiate between embryogenic callus consisting of somatic embryos and non-embryogenic callus. In Musa spp., globular to torpedo shaped somatic embryos resulted in good suspension cultures followed by recovery of somatic embryos during developmental stage. The morphological observations were supported by histological sections of structures at all particular stages. Histological examination also revealed the vascular connection in germinated somatic embryos which later turned into a complete plantlet. 相似文献
14.
T. R Monteiro E. O Freitas G. F Nogueira 《The Journal of Horticultural Science and Biotechnology》2018,93(2):196-203
The current analysis describes an improved protocol for somatic embryogenesis and plant regeneration in oil palm (Elaeis guineensis) through liquid medium, and assesses the influence of successive subcultures during induction of calluses in three Brazilian oil palm varieties. Calluses were induced in a Murashige and Skoog (MS) medium with 450 Picloram, 0.5 g L?1 glutamine, 2.5 g L?1 activated charcoal, 30 g L?1 sucrose, and solidified with 2.5 g L?1 Phytagel. In a first experiment, the effect of continued subculture of explants every 30 days to fresh culture medium was determined. During a second experiment, part of the embryogenic calluses obtained were transferred to a liquid medium under agitation, consisting of MS with 5 µM picloram or 2,4-dichlorophenoxyacetic acid (2,4-D). After 210 days, the calluses were transferred to semi-solid media for differentiating somatic embryos. It was observed that continued subculture of explants monthly was a determinant in stimulating and improving the formation of embryogenic calluses. Embryogenic calluses in liquid medium with 2,4-D significantly improved the percentage of differentiated somatic embryos (up to 80.2%), with the largest amount of torpedo embryos (8.3 per callus). Regenerated plants with roots were individualised and transferred to a greenhouse, with close to 95% survival. 相似文献
15.
以山核桃（Carya cathayensis Sarg.）自然授粉后10周的幼胚为外植体,对影响胚性愈伤组织和体胚发生的主导因子（基本培养基、植物生长调节物质等）进行了比较分析;对影响体胚萌发的脱水处理时间进行了比较;并采用石蜡切片法对幼胚脱分化产生胚性愈伤组织及体胚发生发育过程进行了组织细胞学观察。结果表明,幼胚胚轴和子叶接种在基本培养基1/2 MS上,胚性愈伤组织诱导率显著高于其它处理,达45.3%。接种于基本培养基DW中的幼胚体细胞胚诱导率显著高于其它培养基处理,达16.7%。显微镜下可观察到球形胚、心形胚、鱼雷胚和子叶胚。培养基添加1.0 mg · L-1 6-BA和0.01 mg · L-1 picloram（氨氯吡啶酸）组合时,胚性愈伤组织诱导率最高,为48.6%;而1.0 mg · L-1 6-BA与0.001 mg · L-1 picloram组合,体胚诱导率最高,为23.6%。体细胞胚发生方式属于间接体胚发生。用饱和Ca(NO3)2 · 4H2O对体胚进行脱水处理,脱水处理3 d后萌发率为39.03%,显著高于对照及其余处理。将脱水处理后的体胚接种至WPM基本培养基中,光照条件下培养,10周后可长成具3 ~ 4片真叶的完整植株。 相似文献
16.
The nucellus and globular adventitious proembryos were removed from 2-month-old fruits of mango (Mangifera indica L.) cultivars ‘Ono’ and ‘Chino’, and were cultured on sterile, solid Murashige and Skoog (MS) medium that had been modified as follows: half-strength major salts and chelated iron; 20% (v/v) coconut water (CW); 6% sucrose; 100 mg l?1 ascorbic acid and 400 mg l?1 glutamine. Embryogenic explants were sub-cultured after 4–6 weeks in liquid modified MS medium containing 2 mg l?1 2,4-dichlorophenoxyacetic acid (2,4-D) instead of CW. Rapidly growing cultures were established and were sub-cultured monthly. Somatic embryogenesis was induced following sub-culture from MS medium with 2,4-D to MS without growth regulators and with or without activated charcoal (0.5%). Germination of somatic embryos appeared to be enhanced by 1 mg l?1 benzyladenine (BA); however, most of the germinating embryos became embryogenic. 相似文献
17.
酿酒葡萄‘神索’体胚发生及再生体系遗传稳定性分析 总被引：7，自引：0，他引：7
以酿酒葡萄‘神索’未成熟合子胚为外植体, 通过植物生长调节剂、光照、温度等因素的控制, 研究了葡萄体胚的产生、保存及植株再生。结果表明, 以NN为基本培养基, 诱导胚性愈伤组织的适宜植物生长调节剂水平是1.0 mg·L - 1 2,4-D; 诱导体胚的生长调节剂水平是1.0 mg·L -1 NAA + 0.5 mg·L - 1 BA, 体胚诱导率为37.5%; 5℃微光条件, 适宜体胚的保存; 体胚成熟及植株再生的植物生长调节剂水平是0.05 mg·L - 1 NAA + 0.5 mg·L - 1 BA, 成苗率为42.1%。利用流式细胞仪并结合染色体计数对体胚及再生植株细胞核DNA含量及染色体鉴定表明, 体胚细胞染色体存在一定的变异, 而体胚再生植株倍性稳定, 细胞核DNA含量及染色体数与供体母株一致。 相似文献
18.
Anandamoy Dam Chayanika Bhattacharya 《The Journal of Horticultural Science and Biotechnology》2017,92(4):411-423
An efficient indirect somatic embryogenesis and Agrobacterium-mediated transformation protocol for Limonium sinense has been established, wherein neomycin phosphotransferase II (npt II) and β-glucuronidase (GUS) genes were used as selectable and screenable markers, respectively. The efficiency of plantlet regeneration from transformed tissue was compared between direct embryogenesis from leaf and indirect embryogenesis from callus. Embryogenic callus (EC) was initiated from leaf explants on MS medium supplemented with 6.7 μM 2,4-D and 2.22 μM BA. The somatic embryos were induced, matured, and germinated when ECs were transferred onto MS medium supplemented with 4.44 μM BA and 1.07 μM NAA. Agrobacterium tumefaciens strain LBA 4404 containing the vector pBI121 was used for the transformation. Transient GUS expression frequency was evaluated and putative transgenic plants were successfully grown on culture medium in presence of kanamycin (80–100 mg L?1). PCR analysis of putative transgenic plants confirmed the presence of GUS and nptII genes. The transformation efficiency obtained through indirect embryogenesis from calluses (4%) was much higher than through direct embryogenesis from leaf explants (0.9%). 相似文献