本文選題：衛(wèi)矛 + 不定芽　； 參考：《沈陽農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：本研究以衛(wèi)矛(Euonymus alatus(Thunb.)Sieb)不同部位為外植體,探討了取材時(shí)間及滅菌方法對(duì)莖段消毒效果的影響;研究了沙藏與未沙藏、成熟與未成熟、帶假種皮與去除假種皮及GA3濃度4種因素對(duì)種子萌發(fā)的影響,沙藏與未沙藏的成熟種胚、NAA(0.5、1.0、2.0mg/L)和 6-BA(0、1.0mg/L)對(duì)不定芽誘導(dǎo)的影響,以及 NAA(0.5、0.05mg/L)、6-BA(1.0、2.0mg/L)和瓊脂(6.5、7.0g/L)對(duì)莖段不定芽誘導(dǎo)的影響,探討了基本培養(yǎng)基、2,4-D、TDZ、暗培養(yǎng)時(shí)間4個(gè)因子對(duì)花生殖器官(花萼、花瓣、雄蕊、雌蕊)和葉片的愈傷組織誘導(dǎo)的影響,從而篩選出最佳外植體及誘導(dǎo)方式;繼而在基本培養(yǎng)基、6-BA、NAA、GA34個(gè)因子3個(gè)水平中選出不定芽增殖的最佳培養(yǎng)基;最終從添加NAA、IAA、IBA(0.2-4.0mg/L)及GA3(0.2-0.8mg/L)的28個(gè)處理中篩選出試管內(nèi)生根最佳方法,并與試管外生根下的基質(zhì)、生根劑、激素濃度的4個(gè)水平中的最佳生根方法進(jìn)行了比較,篩選出衛(wèi)矛最佳生根法。研究結(jié)果如下:(1)莖段最佳取材時(shí)間是5月中旬,染菌率為3%,存活率76.67%;最佳消毒方法是Hgc12 8min并加入兩滴吐溫-20,存活率59.67%。(2)去假種皮的沙藏種子在MS+GA32.0mg/L培養(yǎng)基中培養(yǎng)20d后萌發(fā)出子葉,種子萌發(fā)誘導(dǎo)率(10.2%)高于其他處理。含葉中脈的葉片在WPM+2,4-D 0.8 mg/l+ TDZ 1.0 mg/1培養(yǎng)基中的愈傷組織誘導(dǎo)率最高(65%-67%),但沒有 B5+2,4-D 1.2 mg/l+ TDZ 2.0 mg/l+暗培養(yǎng) 20d處理中的雌蕊愈傷組織誘導(dǎo)率(均值86.5%)高。雌蕊愈傷組織為淺綠疏松狀,并不是最佳愈傷組織形態(tài)。因而用衛(wèi)矛葉片、雌蕊作為外植體誘導(dǎo)愈傷組織未取得成功,還有待研究。種胚不定芽誘導(dǎo)的最佳培養(yǎng)基為MS+6-BA1.0mg/L +NAA0.5mg/L,不定芽誘導(dǎo)率為12.8%。而莖段在6-BA2.0mg/L+NAA0.05mg/L培養(yǎng)基中的不定芽誘導(dǎo)率可達(dá)70.67%,遠(yuǎn)遠(yuǎn)高于種胚,因而莖段可為衛(wèi)矛不定芽誘導(dǎo)的最佳外植體。(3)MS+6-BA1.0mg/L+NAA0.05mg/L是衛(wèi)矛不定芽增殖的最佳培養(yǎng)基,增殖系數(shù)為23.51,即獲得的叢生芽芽數(shù)為23.51個(gè)。在其上繼代培養(yǎng)3次后,叢生芽芽數(shù)可增殖到48.5個(gè)。(4)衛(wèi)矛試管內(nèi)生根最佳培養(yǎng)基為1/2MS+GA30.4mg/L,生根率72.5%,平均生根數(shù)4.1條。試管外生根的最佳條件是蛭石+IAA2500mg/L,生根率為100%,平均生根數(shù)為5.3條。兩種生根方法均可有效促使衛(wèi)矛組培苗生根,但以試管外生根更佳。
[Abstract]:In this study, the different parts of Euonymus alatusus Thunb. Sieb were used as explants to investigate the effects of sampling time and sterilization methods on the sterilizing effect of stem segments, and to study the effects of sand storage and non-sand storage, mature and immature. The effects of four factors on seed germination, such as seed coat removal and seed coat removal and GA3 concentration, the effects of sand storage and non-sand storage on adventitious bud induction, and the effects of sand storage on adventitious bud induction, and the effects of sand storage and non-sand storage on adventitious bud induction, and the effects of NAA 0.55 mg / L 6-BA1.02.0 mg / L) and Agar 6.5g / L on adventitious bud induction. The effects of four factors on the callus induction of floral reproductive organs (calyx, petals, stamens, pistil) and leaves were studied. Then, the best medium for adventitious bud proliferation was selected from 3 levels of 34 factors of basic medium 6-BAANAA, and the best method of rooting in vitro was screened out of 28 treatments supplemented with NAA AIAA IBA 0.2-4.0 mg / L and GA _ 3N _ 0.2-0.8 mg / L), and the best rooting medium, rooting agent, was found with the substrate under rooting outside the tube, rooting agent. The best rooting method was screened out from the four levels of hormone concentration. The results were as follows: (1) the best sampling time of stem segment was in mid-May, the rate of bacterial infection was 3 and the survival rate was 76.67.The best disinfection method was Hgc12 8min and two drops of Tween -20, the survival rate was 59.67.2.) the seeds of sand-covered seed were cultured in MS GA32.0mg/L medium for 20 days and germinated to produce cotyledons. Seed germination induction rate was higher than other treatments. The callus induction rate of leaves with midvein was higher than that of B5224-D 1.2 mg/l TDZ 2.0 mg/l in WPM 24-D 0.8 mg/l TDZ 1.0 mg/1 medium, but the callus induction rate (mean 86.5) was higher than that of B5224-D 1.2 mg/l TDZ 2.0 mg/l dark culture for 20 days. The pistil callus is light green and loose, and is not the best callus morphology. Therefore, the callus induction with the leaves and pistil as explants has not been successfully induced, which remains to be studied. The best medium for adventitious bud induction was MS 6-BA1.0mg/L NAA 0.5 mg / L, and the rate of adventitious bud induction was 12. 8%. The adventitious bud induction rate of stem segment in 6-BA2.0mg/L NAA0.05mg/L medium was 70.67, which was much higher than that of seed embryo. Therefore, stem segment could be the best explant for adventitious bud induction of Leptanthus mongolicus L. 6-BA1.0mg/L NAA0.05mg/L was the best medium for adventitious bud proliferation. The multiplication coefficient was 23.51, that is, the number of buds was 23.51. After 3 times of subculture, the number of buds could proliferate to 48.5%. The best medium for rooting in vitro was 1/2MS GA30.4mg / L, the rooting rate was 72.5% and the average number of rooting was 4.1. The optimum conditions for rooting in vitro were vermiculite IAA 2500 mg / L, rooting rate was 100 and the average number of rooting was 5.3. The two rooting methods can effectively promote the rooting of the plantlets in vitro, but it is better to take root in vitro.
【學(xué)位授予單位】：沈陽農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S687
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本文編號(hào)：1792515