| 木薯种子体细胞胚诱导发生及植株再生体系建立 |
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| 引用本文： | 韦祖生,杨秀娟,付海天,田益农.木薯种子体细胞胚诱导发生及植株再生体系建立[J].南方农业学报,2017,48(12). |
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| 作者姓名： | 韦祖生 杨秀娟 付海天 田益农 |
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| 作者单位： | 1. 广西亚热带作物研究所,南宁530001;广西木薯研究所,南宁530001;2. 广西亚热带作物研究所,南宁530001;农业部亚热带果品蔬菜质量监督检验测试中心,南宁530001 |
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| 基金项目： | 国家木薯产业技术体系建设项目，广西自然科学基金项目，广西科学研究与技术开发计划项目 |
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| 摘 要： | 【目的】建立木薯杂交种子诱导体细胞胚发生及植株再生体系,为木薯育种及种质创新提供技术参考。【方法】以不同成熟度的木薯华南5号(SC5)杂交种子组织为外植体材料,包括未成熟种子子叶(a1)和胚芽(b1)、成熟种子子叶(a2)和胚芽(b2)及诱导萌发种子子叶(a3)、胚芽(b3)和胚轴(c3),利用不同外源激素诱导其体细胞胚胎发生、子叶器官形成、不定芽(茎叶器官)发生和生根,并比较不同外源激素的诱导效果。【结果】利用基本培养基(M1)预培养外植体材料可有效控制其污染率(5.00%),保持其生物活性(90.00%),预培养材料的褐化率比对照(未预培养,CK)降低30.00%~40.00%,且预培养的子叶(a)、胚芽(b)和胚轴(c)膨大率分别比CK提高6.67%~13.33%、6.67%~50.00%和20.00%。利用4.00 mg/L 2,4-D和12.00 mg/L Picloram诱导体细胞胚胎发生,结果发现胚芽出胚率较高,胚轴次之,子叶较低;成熟度高的种子材料出胚率高于成熟度低的种子材料;4.00 mg/L 2,4-D诱导的体细胞胚胎发生效果优于12.00 mg/L Picloram。0.10 mg/L BAP和1.00 mg/L NAA均可诱导体细胞胚胎分化、增殖,但0.05 mg/L BAP+0.50 mg/L NAA诱导下a、b和c体细胞胚胎增殖率均比单独添加0.10 mg/L BAP或1.00 mg/L NAA显著提高(P0.05,下同)。a2、a3、b1、b2、b3和c3的不定芽发生率分别为30.00%、40.00%、40.00%、53.33%、70.00%和13.33%,即对于同一组织,成熟度高的种子材料体细胞诱导不定芽发生效果优于成熟度低的种子材料,且不同成熟度的种子均以b最优。a2、a3、b1、b2、b3和c3的生根率分别为23.33%、23.33%、16.67%、30.00%、56.67%和0,其中b3的生根率显著高于其他材料,a2、a3、b1和b2的生根率无显著差异,也表现为成熟度高的种子材料比成熟度低的种子材料更易诱导生根,发育更好。7种不同成熟度的杂交种子材料组织中,a2、a3、b1、b2和b3经体细胞诱导获得再生植株,成功率为71.43%。【结论】通过预培养可有效防止木薯杂交种子组织褐化,提高其膨大诱导效果。在木薯生产中,应在培养基中添加4.00 mg/L2,4-D进行体细胞胚胎发生诱导,混合添加0.05 mg/L BAP和0.50 mg/L NAA进行体细胞胚胎增殖培养,且应选择成熟度高的种子材料(如萌发种子胚芽等)作为外植体材料。
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| 关 键 词： | 木薯 种子 体细胞 诱导 再生 |
Induction of somatic embryogensis and plant regeneration system in cassava seed |
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| WEI Zu-sheng,YANG Xiu-juan,FU Hai-tian,TIAN Yi-nong.Induction of somatic embryogensis and plant regeneration system in cassava seed[J].Journal of Southern Agriculture,2017,48(12). |
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| Authors: | WEI Zu-sheng YANG Xiu-juan FU Hai-tian TIAN Yi-nong |
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| Abstract: | Objective]Somatic embryogensis induction of cassava hybrid seed and seedling regeneration system were constructed to provide technology reference for cassava breeding and germplasm innovation.Method]Hybrid seeds of cassava variety Southern China 5(SC5)at various maturity degrees were used as explant materials,including cotyledons (a1)and germs(b1)of immature seed,cotyledon(a2)and germ(b2)of mature seed,cotyledon(a3),germ(b3)and hy-pocotyl(c3)of germinated seed.Different exogenous hormones were used to induce the somatic embryogenesis,forma-tion of cotyledon organ,development and rooting of adventitious bud(stem and leaf organ).And the induction effects of these exogenous hormones were compared.Result]Pre-culture with basic culture medium(M1)could control the pollu-tion rate(<5.00%)of explants,maintain their biological activities(>90.00%).The browning rate of pretreated materials decreased 30.00%-40.00% than control(not pretreated,CK),and swelling rates of cotyledon(a),germ(b)and hypocotyl (c)increased 6.67%-13.33%,6.67%-50.00% and 20.00% respectively.Inducing somatic embryogenesis with 4.00 mg/L 2,4-D and 12.00 mg/L Picloram,somatic embryogenesis induction rate of germ was the highest,hypocotyl was the se-cond and cotyledons was the lowest.Somatic embryogenesis induction rate of mature seed material was higher than imma-ture seed material,and the effects of 4.00 mg/L 2,4-D was better than that of 12.00 mg/L Picloram.0.10 mg/L BAP and 1.00 mg/L NAA could induce differentiation and proliferation of somatic embryogenesis.But the induction rates of soma-tic embryogenesis in a,b and c induced by 0.05 mg/L BAP+0.50 mg/L NAA were significantly(P<0.05,the same below) increased compared with those adding single 0.10 mg/L BAP or 1.00 mg/L NAA.The adventitious buds induction rate were 30.00%(a2),40.00%(a3),40.00%(b1),53.33%(b2),70.00%(b3)and 13.33%(c3)respectively,which showed that for the same tissue,somatic embryogenesis induction rate of mature seed materials were better than immature seed materials.Among all the seed materials with various maturities,the effects of b were the best. The rooting rates were 23.33%(a2),23.33%(a3),16.67%(b1),30.00%(b2),56.67%(b3)and 0(c3)respectively;the rooting rate of b3 was significantly higher than other materials,and there was no significant difference between a2,a3,b1 and b2.It showed that for the same tissue,rooting induction of mature seed materials was easier than immature seed materials,and their de-velopment was also better.Out of seven hybrid seed materials with various maturities,regeneration plants of a2,a3,b1, b2 and b3 were obtained through somatic embryogenesis induction with success rate of 71.43%.Conclusion]Pre-culture can inhibit browning of cassava hybrid seed tissues and increase the swelling induction effects. In cassava planting, adding 4.00 mg/L 2,4-D in medium can induce somatic embryogenesis,and adding 0.05 mg/L BAP and 0.50 mg/L NAA misture in medium can proliferate somatic embryogenesis.In addition,seed materials with high maturity level(e.g.em-bryo of germinated seed)should be selected as explant materials. |
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| Keywords: | cassava(Manihot esulenta Crantz) seed somatic cell induction regeneration |
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