【摘要】：本試驗(yàn)分別以沙地云杉未成熟合子胚與成熟合子胚為外植體,研究不同基本培養(yǎng)基、不同激素組合,不同株系對(duì)胚性愈傷誘導(dǎo)、增殖以及體胚分化成熟的影響,建立并優(yōu)化了沙地云杉體細(xì)胞胚胎發(fā)生體系,結(jié)果表明:1.沙地云杉胚性愈傷組織的誘導(dǎo):以未成熟合子胚為外植體誘導(dǎo)胚性愈傷組織最適誘導(dǎo)條件包括:最佳的激素組合為添加6-BA 1.2～1.8 mg/L、2,4-D 1.8～2.7 mg/L,胚性愈傷組織誘導(dǎo)率最高可達(dá)74.00%;最佳采集日期為8月5日前后,胚性愈傷組織誘導(dǎo)率最高為67.70%;不同株系胚性愈傷組織誘導(dǎo)率最低為15.91%,最高可達(dá)77.50%,且誘導(dǎo)率與其飽籽率呈顯著正相關(guān)(r=0.865,P=0.006)關(guān)系;以成熟合子胚為外植體誘導(dǎo)胚性愈傷組織的培養(yǎng)條件為:初代培養(yǎng)采用LM +6-BA 2.0 mg/L + KT 1.0 mg/L + 2,4-D 4.0 mg/L,繼代培養(yǎng)采用 1/2LM + 6-BA 1.0 mg/L + KT 0.5 mg/L + 2,4-D 2 mg/L,胚性愈傷組織誘導(dǎo)率為12.00%,其它處理沒(méi)能誘導(dǎo)產(chǎn)生胚性愈傷組織。2.胚性愈傷組織增殖最佳激素組合為添加6-BA 0.8 mg/L、2,4-D 0.8～1.6 mg/L,在愈傷組織增殖過(guò)程中,2,4-D的作用更明顯,最高增殖率為7.24倍。3.沙地云杉體細(xì)胞胚胎分化與成熟最適培養(yǎng)條件為:1/2LM+ABA 22.5mg/L +PEG4000 45 g/L,體胚分化數(shù)最高為362個(gè)/g,畸形胚數(shù)為108個(gè)/g,畸形胚率為29.83%。K-81株系體胚分化數(shù)最高為573.4個(gè)/g,其中畸形胚數(shù)為151個(gè)/g,畸形胚率為26.33%。4.沙地云杉體細(xì)胞胚胎發(fā)生發(fā)育過(guò)程組織細(xì)胞學(xué)觀察表明:胚性愈傷組織是由橢圓形或長(zhǎng)條形的胚性細(xì)胞以及由胚性細(xì)胞分裂所產(chǎn)生的胚性胚柄團(tuán)組成;在愈傷組織成熟發(fā)育過(guò)程中,體細(xì)胞胚胎發(fā)育共經(jīng)過(guò)早期原胚、球形胚,早期體胚及子葉胚四個(gè)階段。
[Abstract]:In this experiment, immature zygotic embryos and mature zygotic embryos of Picea spruce were used as explants to study the effects of different basic media, different hormone combinations and different strains on embryogenic callus induction, proliferation and somatic embryo differentiation and maturation. The somatic embryogenesis system of Picea spruce was established and optimized. Embryogenic callus induction of Picea spruce: the optimum conditions for embryogenic callus induction with immature zygotic embryos as explants are as follows: the best hormone combination is the rate of embryogenic callus induction added with 6-BA 1.2n 1.8 mg/L,2,4-D 1.82.7 mg/L,. Up to 74.00; the best date of collection is around August 5, The highest rate of embryogenic callus induction was 67.70%, the lowest rate of embryogenic callus was 15.91, the highest was 77.50%, and there was a significant positive correlation between the induction rate and the full seed rate (r 0.865P0. 006). The culture conditions of embryogenic callus induced by mature zygote embryos were as follows: the primary culture was carried out with LM 6-BA 2.0 mg/L KT 1.0 mg/L 2G 4-D 4.0 mg/L, subculture and 1/2LM 6-BA 1.0 mg/L KT 0.5 mg/L 2N 4-D 2 mg/L, embryos. The rate of callus induction was 12.00%, while other treatments failed to induce embryogenic callus. The best hormone combination of embryogenic callus proliferation was that the addition of 6-BA 0.8 mg/L,2,4-D 0.81. 6 mg/L, had a more obvious effect on callus proliferation, and the highest proliferation rate was 7.24 times. 3. The optimum culture conditions for somatic embryo differentiation and maturation of Picea spruce are as follows: 1 / 2 LM ABA 22.5mg/L PEG4000 45 g / L, somatic embryo differentiation highest 362 / g, abnormal embryo number 108 / g, abnormal embryo rate of 29.83%.K-81 strain highest 573.4% / g. The number of embryos is 151 / g, and the rate of abnormal embryo is 26.33. 4. Histological observation of somatic embryogenesis and development of Picea spruce showed that embryogenic callus was composed of elliptical or long embryonic cells and embryogenic petiole produced by embryogenic cell division. During the process of callus maturation, somatic embryogenesis went through four stages: early proembryo, globular embryo, early somatic embryo and cotyledon embryo.
【學(xué)位授予單位】：內(nèi)蒙古農(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)】：S791.18

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