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正交试验法在龙眼和荔枝组织培养中的应用 总被引：13，自引：0，他引：13
利用正交设计试验法研究了椰乳（ＣＭ）、２,４－Ｄ、赤霉素（ＧＡ３）和蔗糖等四个因素及其组合对石硖龙眼和桂味荔枝的幼胚愈伤组织产生的影响,并讨论了正交设计试验法应用于龙眼和荔枝组织培养基筛选的可能性。结果表明：１）对龙眼,诱导愈伤组织的最优组合是ＣＭ１０％、２,４－Ｄ１．０ｍｇ．ｌ＾－１和蔗糖３％;对荔枝,最优组合是ＣＭ０％,２,４－Ｄ１．０ｍｇ．ｌ＾－１,ＧＡ３０ｍｇ．ｌ＾－１和蔗糖５％。２）适当 相似文献
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苹果砧木M26离体叶片愈伤组织发生及分类研究 总被引：1，自引：0，他引：1
对苹果砧木Ｍ２６ 离体叶片愈伤组织的发生研究表明,取继代后３０ ～３５ ｄ 的试管苗顶部１～４ 叶位的叶片,沿与主脉垂直方向横切后,以叶背面朝上方式置于愈伤组织诱导培养基上,经观察在叶片近柄端切口处愈伤组织发生较早,产生量较多。最适的愈伤组织诱导培养基为：ＭＳ＋２ ,４ － Ｄ０ ．２ ～２ ．０ ｍｇ·ｌ－ １ ＋ＩＡＡ０ ．２ ～２．０ ｍｇ·ｌ－ １ ＋ ＢＡ０ ．５ ～１．０ ｍｇ·ｌ－ １ ＋ 蔗糖３ ％ ＋ 琼脂０．６％ 。将诱导产生的愈伤组织按外部形态、细胞特点及生理生化指标等分为三类,其中表面干燥致密、细胞内含物丰富、淀粉、蛋白质及干物质含量均较高的Ⅲ类愈伤组织的芽再生频率最高,可达８３ ．３３ ％ ,是能利用的愈伤组织类型。 相似文献
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通过以甜菜未成熟胚为外植体诱导愈伤组织，再将产生的愈伤组织转移至分化培养基（ＭＳＢ培养基附加１．０ｍｇ／Ｌ６－ＢＡ，３％蔗糖，８％琼脂）进行分化力测试实验，筛选出两个再生能力强的基因型－８９３０，ＧＤ２。通过挑选淡黄色或灰白色，呈颗粒状的愈组织继代培养，经调节培养基的ＩＡＡ和ＮＡＡ的浓度，得到再生能力强的胚性愈伤组织，建立了胚性细胞无性系。 相似文献
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酿酒葡萄原生质体再生植株 总被引：1，自引：0，他引：1
将酿酒葡萄品种白诗南、梅郁的花丝接种在含６ＢＡ２．０ｍｇ·１－１、２,４－Ｄ０．５ｍｇ·１－１的Ｂ５诱导培养基上,诱导产生胚性愈伤组织。胚性愈伤组织经液体悬浮培养形成含大量胚性细胞团的悬浮培养物。用Ｃｅｌｌｕｌａｓｅ Ｒｓ２％、Ｐｅｃｔｏｌｙａｓｅ Ｙ２３０．３％,５ｍｍｏｌ·１－１ＣａＣｌ２·２Ｈ２Ｏ、０．６ｍｏｌ·１－１甘露醇、０．３％葡聚糖硫酸钾、ｐＨ５．６～５．８的酶混合液从胚性细胞团分离得到原生质体。原生质体培养到第５天出现第一次分裂,４０ｄ形成上百个细胞的大细胞团,５０ｄ形成０．５～１．０ｍｍ大小的小愈伤组织。这些小愈伤组织在含６ＢＡ０．５ｍｇ·１－１的Ｂ５分化培养基上分化出胚状体进而形成幼苗,在生根培养基上生根形成再生植株。 相似文献
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猕猴桃的组织培养和快速繁殖 总被引：4，自引：0，他引：4
１ 植物名称 金魁猕猴桃（Ａｃｔｉｎｉｄｉａ ｄｅｌｉｃｉｏｓａｖａｒ．Ｊｉｎ Ｋｕｉ）２ 材料类别 嫩茎３ 培养条件 培养基：①ＭＳ＋ＺＴＷ．０ｍｓ／Ｌ；②ＭＳ＋ＺＴ１．０ｍｐ／Ｌ；＠ ＭＳ＋６ＢＡ２．０ｍｇ／Ｌ＋ＮＡＡ ０．１ｍｇ／Ｌ；④１／２ＭＳ＋ＩＢＡ０．５～０．７ｍｇ／Ｌ。 蔗糖（市售白砂糖）浓度①～③培养基为３．０％，④培养基为 ２．０％；琼脂浓度为 ０．６５％～０．７０％，ｐＨ５．８～６．０，培养温度 ２５～２８℃，每天光照 １３ｈ，光照强度 １５００～２０００ １Ｘ。４ 生长与分化情况４．１ 愈伤… 相似文献
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银杏成熟胚乳培养的细胞组织学观察 总被引：9，自引：0，他引：9
诱导银杏成熟胚乳产生愈伤组织时无需胚的存在，单独使用生长素或在细胞分裂素／生长素为０．５－０．９的范围内均能诱导部分胚乳切块产生愈伤组织。并且在Ｗｈｉｔｅ ＋Ｋｔ２．７ｍｇ．１＾＿１＋２．４－Ｄ１．０ｍｇ．ｌ＾－１＋ＮＡＡ２．０ｍｇ．ｌ＾－１的培养基上，愈伤组织的诱导率最高，达到８３．３％。胚乳细胞脱分化进行有丝分裂和无丝分裂，其中无丝分裂可以观察到劈裂和缢裂两种类型。愈伤组织起源于胚乳表皮及其下 相似文献
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培养基及培养条件对银杏愈伤组织黄酮产量的影响 总被引：34，自引：1，他引：33
在固体培养条件下，研究了不同外植体来源、培养基、抗褐变剂及培养时间等因子对银杏愈伤组织黄酮产量的影响。结果表明：①不同外植体来源的愈伤组织黄酮含量为叶片＞茎段＞子叶；②光照培养的愈伤组织黄酮含量显著高于暗培养愈伤组织黄酮含量；③愈伤组织培养４０～４５ｄ，黄酮含量接近最大值；④Ｗｈｉｔｅ培养基上的愈伤组织黄酮含量显著高于ＭＴ培养基，但黄酮生物产量却相反；⑤不同激素组合中，以ＢＡ与ＮＡＡ组合效果最好；⑥在ＰＡ（植酸）、ＰＶＰ（聚乙烯吡咯烷酮）、ＰＭ（ＰＶＰ＋ＭＥＳ）、ＡＣ（活性炭）和维生素Ｃ等５种抗褐变剂中，以ＰＭ效果最好。黄酮生产最佳培养条件为：叶片愈伤组织在ＭＴ＋ＢＡ１．０ｍｇ／Ｌ＋ＮＡＡ３．０ｍｇ／Ｌ＋糖５％＋ＰＭ（ＰＶＰ２．０％＋ＭＥＳ０．０５％）培养基上光照培养４０～４５ｄ。 相似文献
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菠萝叶基愈伤组织诱导体细胞胚 总被引：6，自引：1，他引：6
以菠萝[Ananascomosus(L.)Merr.]吸芽上的叶基部组织为材料,研究了2,4-D、NAA、BA、TDZ在菠萝愈伤组织诱导体细胞胚过程中的影响。结果表明,2,4-D在体细胞胚诱导过程中起着极为重要的作用,其适宜浓度为5mg/L,适当浓度(1mg/L)的BA在降低畸形胚比例和增加体细胞胚发生率方面有重要作用,但BA也会促进愈伤组织分化不定芽;低浓度的TDZ与BA效果相似,在不同生长调节物质处理组合中,MS+0.01mg/LTDZ+5mg/L2,4-D培养基的体细胞胚发生率最高(97%)。低浓度NAA(2￣5mg/L)利于非胚性愈伤组织的增殖,而高浓度的NAA虽能促进体细胞胚发生,但这种体细胞胚很难继续增殖,在萌发时只生根不发芽。成熟体细胞胚在MS+0.2%活性炭上30d时的转株率为22.7%。 相似文献
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Effect of sucrose concentrations on somatic embryogenesis in carnation (Dianthus caryophyllus L.) 总被引：1，自引：0，他引：1
Omid Karami Ali Deljou Mahmoud Esna-Ashari Prisa Ostad-Ahmadi 《Scientia Horticulturae》2006,110(4):340-344
The effect of sucrose concentration on callus induction followed by differentiation of embryogenic callus derived from petal explants of four carnation cultivars (Nelson, Sagres, Spirit and Impulse) was investigated. Embryogenic calli were produced on Murashige and Skoog [Murashige, T., Skoog, F.A., 1962. Revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant 154, 73–479] basal medium (MS) culture medium containing six concentrations of sucrose (3, 6, 9, 12, 15 and 18%, w/v) all supplemented with 9 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.8 μM 6-benzyladenine (BA). Maximum frequency of embryogenic callus was obtained from the media containing 9 and 12% sucrose. Somatic embryos were induced on a hormone-free MS media containing the seven concentrations of sucrose. Development of somatic embryos was enhanced by increasing sucrose concentration from 1.5 to 12%, while it was reduced in higher concentrations of 15 and 18%. However, normal embryos were not developed in the media containing 1.5 and 3% sucrose. Ninety-five percent of somatic embryos were regenerated to form the entire plantlets when they transferred onto the half-strength hormone-free MS culture medium containing 3% sucrose. Plantlets were also continued to grow normally under greenhouse condition. 相似文献
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Akhtar Nasim 《The Journal of Horticultural Science and Biotechnology》2013,88(6):556-562
SummaryAn embryogenic protocol for plant regeneration of guava (Psidium guajava L.) was established using 10-week post-anthesis, zygotic embryo explants. Somatic embryogenesis was induced on Murashige and Skoog medium (MS) containing 3% (w/v) sucrose, 0.8% (w/v) agar and various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) by continuous treatment of the zygotic embryo explants. Somatic embryos appeared as globular structures at the end of the third week from culture initiation, and heart-shaped, cotyledonary-stage, and torpedo-stage embryos appeared within the next few weeks. The development of somatic embryos was asynchronous and showed five-to-seven discernible stages. Depending upon the response of the somatic embryos during their maturation, germination, acclimatisation, and encapsulation, they were grouped into one of three categories. The preferred type of somatic embryos (≥ 1.5 mm) were called the “elongated torpedo” (ET) category. The slightly less-preferred type of stomatic embryos (from 1.0 – 1.5 mm) were termed the “short torpedo” (ST) category. The least preferred types of somatic embryos, at the cotyledonary, heart-shaped, and/or globular stages of development (< 1.0 mm), were grouped into a third category designated “CHG”. The suitability and efficacy of various growth regulators and other treatments were assessed based on six different embryogenic parameters: (i) the frequency of embryogenesis; (ii) the intensity of embryogenesis, defined as the average number of somatic embryos produced per culture (“ANEPC”); (iii) the frequency of ET somatic embryos; (iv) the frequency of ST somatic embryos; (v) the frequency of CHG somatic embryos; and (vi) the overall efficiency of embryogenesis, defined as the potential of a treament to produce somatic embryos at the ET or ST stages, or at both stages of development, that could be converted into plantlets. In the present report, we found that 0.01 mg l–1 2,4-D gave the maximum frequency and intensity of embryogenesis. But the highest frequencies of ET and ST somatic embryos were produced on MS medium containing 3% (w/v) sucrose and 0.001 mg l–1 2,4-D, while CHG embryos were produced at the highest frequency on the same medium, but containing 0.5 mg l–1 2,4-D. It was difficult to calculate the most effective concentration of 2,4-D for somatic embryogenesis based on parameters (i) – (v) above. Hence, quantitative estimations of the efficiency of embryogenesis (sixth parameter) were imperative in order to analyse the potential of the different treatments. The highest efficiency of somatic embryogenesis was achieved by continuous treatment of 10-week post-anthesis, zygotic embryo explants with 0.01 mg l–1 2,4-D on full-strength MS agar medium containing 3% (w/v) sucrose. These somatic embryos matured normally on the same medium, and germinated well both on half-strength solid and in half-strength liquid MS medium containing 3% (w/v) sucrose. They grew in full-strength liquid MS medium with 3% (w/v) sucrose and showed maximum survival upon transfer to soil and hardening. Evaluations of the efficiency of somatic embryogenesis in guava, based on the six parameters defined above, have also helped us to understand and evaluate processes for high efficiency micropropagation in other species. 相似文献
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以欧洲葡萄‘无核白’（Vitis vinifera‘Thompson Seedless’）未开放的小花蕾为外植体,研究0、1.0、2.0、3.0 mg ? L-1褪黑素（Melatonin）对其体细胞胚的诱导效果。结果表明：‘无核白’小花蕾在 MS + 1.0 ~ 3.0 mg ? L-1 褪黑素 + 2.0 mg ? L-1 6-BA + 30 g ? L-1蔗糖 + 3 g ? L-1植物凝胶的培养基上继代培养30 d时愈伤组织诱导率较好,为73.67% ~ 89.10%,显著高于2,4-D处理,愈伤组织形成所用时间较2,4-D处理缩短了14 d。愈伤组织诱导120 d时,不同浓度褪黑素均出现体细胞胚,其中,以MS + 1.0 mg ? L-1褪黑素 + 2.0 mg ? L-1 6-BA + 30 g ? L-1蔗糖的培养基体细胞胚的发生率最高,180 d时达12.05%。体胚在 MS + 60 g ? L-1 蔗糖 + 0.5 g ? L-1活性炭的X6培养基中萌发30 d后,将萌发的子叶胚转移至MS + 0.2 mg ? L-1 6-BA + 0.1 mg ? L-1 NOA + 30 g ? L-1 蔗糖 + 0.5 g ? L-1活性炭的成苗培养基上,其中,1.0 mg ? L-1褪黑素诱导产生的体胚发育正常的数量较多,为14.84%。不同浓度褪黑素处理的体胚诱导率均于180 d后增长较快,且低浓度的褪黑素有利于体胚萌发与正常发育,2,4-D诱导的愈伤组织无体胚形成。 相似文献
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多因子正交试验对鞭炮花丛生芽诱导条件的筛选 总被引：1，自引：1，他引：0
利用正交试验设计方法探讨6-BA、NAA、2,4-D和蔗糖对丛生芽诱导影响.结果表明:蔗糖和6-BA的浓度对鞭炮花丛生芽诱导的影响最大,高浓度的2,4-D对于愈伤组织的诱导和增殖具有良好的促进作用,但对芽的分化有抑制作用.筛选出诱导鞭炮花丛生芽的最适培养基为MS+2.0 mg/L 6-BA+1.4 mg/L NAA+25 g/L蔗糖+0.7%琼脂.该培养基可使外植体一步诱导出苗,并获得移栽成活的植株. 相似文献
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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
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Meena Joshi Sumit Manjkhola 《The Journal of Horticultural Science and Biotechnology》2013,88(4):570-574
SummaryThis paper reports, for the first time, somatic embryogenesis and synthetic seed production in Selinum tenuifolium Wall. Mature leaf explants inoculated in Murishige and Skoog (MS) medium supplemented with 3 µM 2,4-dichlorophenoxyacetic acid (2,4-D), containing 3% (w/v) sucrose and 0.7% (w/v) agar, induced 67% callus. Maximum production of globular structures, their differentiation into embryos and germination, occurred with a combination of 2 µM benzyladenine (BA) and 2 µM indole-3-butyric acid (IBA). To protect somatic embryos and produce synthetic seeds, gel capsules were standardised using a combination of sodium alginate and calcium nitrate concentrations. Gel capsules were most effective when formed with a combination of 3% (w/v) sodium alginate and 100 mM calcium nitrate for 30 min. The addition of MS medium to alginate capsules with 3% (w/v) sodium alginate, 3% (w/v) sucrose, 2 µM BA and 2 µM IBA significantly improved their germination rate to 77.8%, as well as their resulting shoot length (5.6 cm) and root length (7.2 cm), compared to controls (57.8%). Most plantlets (66%) survived under nursery condition. Storage at 4°C for different periods (10 d or 20 d) significantly (P < 0.05) reduced the percentage survival and germination of somatic embryos and artificial seeds compared to controls or 5 d storage. 相似文献
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荔枝龙眼细胞悬浮培养和转基因研究(英文) 总被引：3，自引：0，他引：3
应用正交设计研究了IBA、6-BA、GA3和CM等植物生长调节剂在龙眼(Dimocarpus longana Lour.)与荔枝 (Litchi chinensis Sonn.)细胞悬浮培养中的作用,并就悬浮培养细胞对Km的敏感性以及用金刚沙对胚性愈伤组织在 液体状态下进行创伤和用分别带有AP1和APBD基因的农杆菌对创伤的愈伤组织进行遗传转化进行了探讨。结果 表明:IBA、6-BA、GA3和CM均可显著地提高悬浮培养系的产量,但GA3导致细胞的快速生长和褐变。悬浮培养系 细胞在液体培养基中比在固体培养基中对Km较敏感。用金刚沙对胚性愈伤组织在液体状态下进行创伤后进行农 杆菌介导转化可以得到更多的Km抗性胚状体。PCR检测结果显示,AP1及AP-D基因均已转入龙眼的胚状体中。 相似文献
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A protocol for plant regeneration by somatic embryogenesis was developed in guava cv. Banarasi local by using immature zygotic embryo explants. Best induction of somatic embryogenesis was achieved from 10-week-old zygotic embryos on MS medium supplemented with 2,4-d (4.52 μM) and 5% sucrose. Maximum number of somatic embryos was produced when zygotic embryo explants were transferred to growth regulator free full strength MS basal medium after 8 days treatment with 2,4-d. Full strength MS basal medium containing 5% sucrose was most favorable for maturation of somatic embryos. Highest frequency of conversion and normal plantlet production were recorded from elongated torpedo stages of somatic embryos on half strength MS medium containing 3% sucrose. Over 90% of rooted shoots survived acclimatization. 相似文献