Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/36733
標題: 黑豆及毛豆未熟子葉培養體胚形成之研究
In Vitro Somatic Embryogenesis of Immature Cotyledon of Black Soybean and Vegetable Soybean
作者: 王瑞章
Wang, Rei-Cheng
關鍵字: Black Soybean;黑豆;Vegetable Soybean;Immature Cotyledon;Somatic Embryogenesis;Embryogenic tissue;毛豆;未熟子葉;體胚形成;胚性組織
出版社: 農藝學系
摘要: 
中文摘要
本研究利用栽培種黑豆台南三號(TN3)及毛豆高雄選一號(KS1)之未成熟種子(長度4-5mm)之子葉為材料,接種於以MSB為基礎培養基,各添加40mg/l 2,4-D及8g/l agar(代號M0a),2g/l gelrite(M0g),以及再各添加10g/l (M1a,M1g),30g/l(M2a,M2g),60g/l sucrose(M3a,M3g)等8種培養基;進行黑豆及毛豆未熟子葉胚性組織與體胚之誘導,並以石臘切片觀察體胚之發育及分化之過程。並將胚性組織團塊繼代於FN Lite培養基,進行胚性懸浮培養試驗。期能建立栽培種黑豆及毛豆之體胚培養技術,以作為基因轉殖研究及遺傳育種時之參考,結果如下︰
1. 黑豆台南三號及毛豆高雄選一號之胚性組織形成率有極顯著差異,黑豆台南三號胚性組織形成率顯著高於毛豆高雄選一號。在不同培養基間,胚性組織形成率亦達極顯著差異,含6%蔗糖及2g/l gelrite 之M3g培養基與含3%蔗糖及2g/l gelrite 之M2g培養基顯著高於其他培養基,未添加蔗糖之M0a及M0g對胚性組織誘導則皆無反應。
2. 胚性組織及球形胚之誘導在培養基與光照處理及品種之交感效應上均呈極顯著,而培養基之組成分的蔗糖及凝膠除在球形胚形成之誘導上與光照處理之交感效應不顯著外,與品種之交感作用及在胚性組織之誘導上均呈極顯著。黑豆台南三號之胚性組織形成率以在含6%蔗糖及2g/l gelrite 之M3g培養基,於暗處理2週後25μmol.m-2s-1 16小時照光6週環境下,其所誘導之胚性組織之形成率71.67%及球形胚46.67%(2.36個)最高,其次為含3%蔗糖及2g/l gelrite 之M2g培養基所誘導胚性組織之形成率33.33%及球形胚23.33%(2.5個)。毛豆高雄選一號則以含3%蔗糖及2g/l gelrite之 M2g培養基,於5-10μmol.m-2s-1 24小時照光8週環境下,誘導胚性組織形成率50%及球形胚36.67%(1.68個)最高。
3. 培養8週後,黑豆台南三號於不同培養基下,其所誘導體胚形態多為胚性組織及球形期胚。毛豆高雄選一號則由含6%蔗糖及8g/l agar 之M3a培養基、含3%蔗糖及2g/l gelrite 之M2g培養基及含6%蔗糖及2g/l gelrite 之M3g培養基誘導結果可看到胚性組織、球形期胚、魚雷期及子葉期胚。
4. 懸浮培養對黑豆及毛豆體胚增殖效果:球形胚增殖倍率黑
豆台南三號為2.8倍,優於毛豆高雄選一號之1.8倍。每個胚性組織平均形成球形胚數黑豆台南三號為3.2個,毛豆KS1則為2.6個。
5.組織學上的觀察,黑豆台南三號及毛豆高雄選一號未熟子葉培養八週後,皆可產生胚性組織及球形期、魚雷期之體胚。黑豆台南三號於培養第七週後開始產生胚性組織,八週後產生球、魚雷期之體胚。

In Vitro Somatic Embryogenesis of Immature Cotyledon of Black Soybean and Vegetable Soybean
Summary
The purposes of the experiments that are to establish a tissue culture system for somatic embryogenesis from immature cotyledon of soybean, and the results will be applied on inducing transgenic plant from explant and embryonic rescue of plant breeding. The explants, immature cotyledons of cv. TN3 of black soybean and cv.KS1 of vegetable soybean, were cultured on modified MSB basal medium containing 2,4-D and other nutrients to induce somatic embryo.
The results are summarized as follows:
1. The formative rates of embryonic tissue of cv. TN3 was significantly higher than it of cv.KS1 . The formative rates of embryonic tissue from immature cotyledons cultured on both M3g and M2g media were significantly higher than others.
2. There were significant interaction between medium and variety on formative rates both embryonic tissue and globular embryo. As to interaction between medium and light was also significantly influential on formation both embryonic tissue and globular embryo.
3. Most of immature cotyledons both cv. TN3 and cv.KS1 cultured in media formated embryonic tissue and globular embryo at 8 weeks olds of explant. However, embryonic tissue, globular embryos and monocotyledonous embryos were only induced with M2g,M3g and M3a media.
4. The formations ratioes of somatic embryos with suspension culture of cv. KS1 and cv. TN3 were 2.8 and 1.8, respectively, and the formations ratios of globular embryos were 2.6 and 3.2, respectively.
5. The differentiation of embryonic tissue from cotyledons of cv. TN3 appeaned at 7 weeks old of explants, and globular embryo and torpedo stage occurred at 8 weeks old of explants.
URI: http://hdl.handle.net/11455/36733
Appears in Collections:農藝學系

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