High Frequency Somatic Embryogenesis and Plant Regeneration of an Elite Chinese Cotton Variety

一個中國棉花優良品種高頻胚胎發生於植株再生

張寶紅(Bao-Hong Zhang) ； 豐嶸(Rong Feng) ； 劉方(Fang Liu) ； 王清連(Qing-Lian Wang)

《Botanical Bulletin of Academia Sinica》 42卷1期 (2001/01) Pp. 9-16

https://doi.org/10.7016/BBAS.200101.0009

摘要

一個中國棉花(Gossypium hirsutum L.)優良品種泗棉3號被選擇作組織培養的材料，選擇獲得了高頻胚胎發生，體細胞胚和再生植株的癒傷組織。在僅附加玉米素的MSB（MS無機鹽和B5維生素）培養基上，三類外植體均誘導獲得了癒傷組織，但癒傷組織的誘導率和誘導量不同。下胚軸切斷比子葉片和根切斷更易誘導獲得癒傷組織。ZT濃度影響棉花癒傷組織的誘導和增值。用於棉花癒傷組織誘導的ZT最佳濃度為3.0~5.0mg/L。接種後連續培養70天可見到胚性和非胚性兩類癒傷組織。當胚性癒傷組織繼代培養到胚分化培養基上培養20天後，可見到不同發育時期的體細胞胚，棉花品種泗棉3號體細胞胚胎發生的能力依賴於外植體的種類和ZT濃度，根最容易誘導獲得胚胎發生，下胚軸次之，子葉最差。低濃度的ZT有利於棉花胚性癒傷組織的誘導。2,4-D的添加雖然有利於癒傷組織的誘導和增殖，但卻不利於體細胞胚的分化、成熟和萌發。活性炭的添加或細胞分裂素ZT和生長素IAA的合理配合使用利於體細胞胚的產生、成熟和萌發。在該實驗中，用於胚性癒傷組織增殖的最佳培養基為附加1.0mg/L 2,4-D,0.5mg/L KT和0.5mg/L ZT的ZH培養基，用於體細胞胚分化和萌發的最佳培養基為附加0.1mg/LZT和2g/L活性炭的MSB培養基。本文建立了一種適用於棉花優良品種泗棉3號的高頻胚胎發生於植株再生的有效方法。利用該方法進行棉花泗棉3號的組織培養，僅需要3~4個月就可以從下胚軸，子葉和根外植體通過胚胎發生途徑誘導獲得完整的再生植株。

關鍵字

陸地棉 ； 泗棉3號 ； 體細胞胚胎發生 ； 植株再生

並列摘要

An elite Chinese cotton (Gossypium hirsutum L.) cultivar Simian-3 was chosen for tissue culture. Callus with a high frequency of somatic embryogenesis, somatic embryos, and regenerative plants was obtained. Callus was induced from three types of explants on MSB (MS salts with B, vitamins) medium supplemented with zeatin (ZT) only, but the percentage of callus induction and growth of callus varied. It appeared that it was much easier to induce callus from hypocotyl than cotyledon or root explants. The concentrations of ZT were critical to the induction and proliferation of callus. The optimum ZT concentration for callus induction was 3.0~5.0mg/L. Two kinds of callus could be identified after 70 days of culture: embryogenic and nonembryogenic callus. Embryogenic callus developed into somatic embryos at various stages after 20 days of subculture. The capability of embryogenesis depended on the explant types. The root was the most responsive explant for production of somatic embryos, the hypocotyl was the next, and the cotyledon was the last. Moreover, a low concentration of ZT was advantageous to the induction of embryogenic callus. 2, 4-dichlorophenoxyacetic acid (2, 4-D) promoted the proliferation of embryogenic callus, but had a negative effect on the differentiation and germination of somatic embryos. Addition of activated charcoal or a proper combination of ZT and 3-indoleacetic acid (IAA) could promote the production, maturation and germination of somatic embryos. The best medium for the proliferation of embryogenic callus was ZH medium (Zhang et al., 1996) with 1.0mg/L 2, 4-D, 0.5mg/L kinetin (KT) and 0.5mg/L ZT. The best medium for the differentiation and germination of somatic embryos was MSB with 0.1mg/L ZT and 2g/L activated charcoal. An efficient protocol for the production of high frequency somatic embryogenesis and plant regeneration of an elite cotton variety Simian-3 has been developed. Complete plants could be regenerated through somatic embryogenesis from hypocotyl, cotyledon and root explants in 3-4 months.

並列關鍵字

Gossypium hirsutum L. ； Plant regeneration ； Simian-3 ； Somatic embryogenesis