發(fā)布時(shí)間：2019-03-30 21:41

【摘要】：實(shí)驗(yàn)通過單因素試驗(yàn)和正交試驗(yàn)的研究方法,優(yōu)化金錢松種子萌發(fā)條件;建立金錢松快速繁育體系;建立金錢松植物細(xì)胞的懸浮培養(yǎng)和植株快速再生的培養(yǎng)體系;探究利用金錢松種子誘導(dǎo)多倍體的最佳培養(yǎng)條件;從金錢松植株的新鮮枝條中分離出內(nèi)生菌,并進(jìn)行了抗菌試驗(yàn)研究。具體實(shí)驗(yàn)結(jié)果如下:(1)通過單因素試驗(yàn)研究了1%多菌靈溶液消毒處理對(duì)其種子的萌發(fā)率和染菌率的影響。實(shí)驗(yàn)結(jié)果表明,隨著1%的多菌靈溶液處理金錢松種子時(shí)間的增長,其種子的萌發(fā)率和染菌率均表現(xiàn)出逐漸下降的趨勢,最有利于提高金錢松種子的萌發(fā)率和降低其種子的染菌率的處理方式為1%多菌靈溶液處理金錢松種子3h,種子的萌發(fā)率為(82±0.32)%,而污染率僅為(3±0.49)%。另外,實(shí)驗(yàn)還研究了不同赤霉素濃度、不同溫度和浸泡不同時(shí)間對(duì)金錢松種子萌發(fā)的影響,研究結(jié)果表明,金錢松種子在60℃下,用6‰赤霉素浸泡2 h后,再放在培養(yǎng)皿中培養(yǎng)30 d,種子萌發(fā)率最高,為97.33%。(2)以金錢松胚軸為外植體誘導(dǎo)愈傷組織的最佳培養(yǎng)基為MS+2,4-D 1.0 mg/L+6-BA 0.5 mg/L+KT 0.1 mg/L;影響胚軸外植體快速產(chǎn)生愈傷組織除了激素配比外,還與外植體的接種方式、切割方式有關(guān);研究培養(yǎng)過程中形成的六種愈傷組織,得出顏色呈綠色、淺綠色的愈傷組織細(xì)胞活力是最高的。(3)金錢松細(xì)胞懸浮培養(yǎng)的最佳培養(yǎng)基MS+6-BA 1.0 mg/L+2,4-D 0.3 mg/L+蔗糖40 g/L+接種量1.2 g/40 ml+p H 6.0;在細(xì)胞的生長過程中,具體表現(xiàn)為,延遲期為0~3 d,指數(shù)期為3~7 d,衰落期為7~20 d;另外,在實(shí)驗(yàn)中觀察還發(fā)現(xiàn),翠綠的愈傷組織生長速度快,黃綠色和黃色的愈傷組織生長速度慢、且褐變速率快。(4)在金錢松植株快速再生研究中,選取金錢松葉片作為外植體誘導(dǎo)愈傷組織,其愈傷組織誘導(dǎo)的最佳培養(yǎng)基為GD+NAA 1.0 mg/L+GA31.0 mg/L,在培養(yǎng)30 d后統(tǒng)計(jì)愈傷組織誘導(dǎo)率為65.4%,誘導(dǎo)不定芽最佳培養(yǎng)基為MS+TDZ 0.2 mg/L+Ad 20 mg/L,培養(yǎng)35 d誘導(dǎo)率為23.1%;誘導(dǎo)不定根的最佳培養(yǎng)基為1/3MS+IAA 1.0 mg/L+活性炭30 mg/L,培養(yǎng)35 d誘導(dǎo)率為54.0%。無菌苗在室內(nèi)環(huán)境下開蓋培養(yǎng)三天以后栽種在大棚中,成活率為93%。(5)在開展金錢松植株的多倍體誘導(dǎo)過程中,最佳誘導(dǎo)培養(yǎng)條件為,在30℃下,0.2%的秋水仙素浸泡36 h,誘導(dǎo)率為78.63%;而誘導(dǎo)率最低的條件為,在20℃下,0.1%的秋水仙素中浸泡12 h,誘導(dǎo)率為49.94%;當(dāng)秋水仙素濃度0.2%,秋水仙素作用時(shí)間從12 h升高到36 h時(shí),金錢松多倍體誘導(dǎo)率逐漸上升,當(dāng)作用時(shí)間從36 h升高為48 h時(shí),誘導(dǎo)率有下降趨勢。(6)本研究從金錢松新鮮枝條中分離出了JJ18、JJ243、JJ314、JJ318、JJ416、JJ27和JJ46共七內(nèi)生菌;用金黃色葡萄球菌為指示菌,篩選出具有抗菌作用的內(nèi)生菌;提取內(nèi)生菌JJ18代謝物,作用于白色念珠菌、紅色毛癬菌、釀酒酵母菌和黑曲霉,測定了各種菌的MIC(最小抑菌濃度)和MBC(最小殺菌濃度)。
[Abstract]:By means of single factor experiment and orthogonal experiment, the germination conditions, rapid propagation system, suspension culture system and plant regeneration culture system of Pinus pinnata were optimized, and the results were as follows: (1) the seed germination conditions of Pinus pines were optimized, and the results were as follows: 1. To explore the optimum culture conditions for inducing polyploidy by using the seeds of Pinus mongolica, endophytic bacteria were isolated from fresh branches of Pinus koraiensis, and the antibacterial test was carried out. The results were as follows: (1) the effect of disinfection treatment with 1% carbendazim solution on the germination rate and infection rate of the seeds was studied by single factor experiment. The results showed that with the increase of 1% carbendazim solution treatment time, the germination rate and the infection rate of the seeds decreased gradually, and the seed germination rate and the infection rate decreased gradually with the increase of the time of 1% carbendazim solution treatment. The best treatment was 1% carbendazim solution for 3 h, the germination rate was (82 鹵0.32)%, and the contamination rate was (3 鹵0.49)%. In addition, the effects of different concentration of gibberellic acid, different temperature and different time on seed germination were studied. The results showed that the seeds were soaked in 6 鈥，

本文編號(hào)：2450514