【作者】 马雯；

【导师】 李唯； 李胜；

【作者基本信息】 甘肃农业大学 ， 发育生物学， 2011， 硕士

【摘要】 大蒜(Allium sativum L.)是一种重要的蔬菜作物,生产中主要靠鳞茎进行无性繁殖。大蒜在长期无性繁殖过程中,易感染和积累多种病毒,且广泛传播,造成种性退化,品质与产量降低。应用大蒜茎尖分生组织并结合热处理是脱除大蒜病毒的行之有效的方法,可以有效脱除大多数病毒,大幅度提高大蒜的品质与产量。但大蒜茎尖培养难度较大,繁殖系数较低,病毒鉴定困难,脱毒良种生产成本较高,成为大蒜脱毒良种培育与生产应用的瓶颈问题。本研究以茎尖分生组织培养结合热处理的方法脱除大蒜病毒,建立大蒜茎尖脱毒培养体系,并以新鲜的或低温贮藏的蒜薹为外植体,研究了贮藏时间和植物生长激素对花苞培养诱导形成气生鳞茎的影响,以及气生鳞茎大小、处理温度和植物激素配比对气生鳞茎的萌发率的影响。利用扫描电镜技术检测大蒜脱病毒前后培养组织所含病毒情况。取得以下研究结果:(1)利用成县迟蒜和鲁蒜王Ⅱ号两个不同品种的鳞茎经行热处理和茎尖脱毒研究,建立了大蒜茎尖脱毒培养体系。结果表明,在MS+0.1 mg/L NAA+0.5 mg/L6-BA的培养基上, 0.3mm左右的大蒜茎尖有较高的萌芽率和试管鳞茎形成率,生长势也较强,萌芽率和试管鳞茎形成率可达到97.4%和87.2%;经试验比较,在37℃下热处理30d后,大蒜茎尖有较高的鳞茎形成率。(2)大蒜花苞可用作离体条件下的外植体材料诱导形成气生鳞茎,但是受到贮藏时间的影响。未经贮藏的外植体在MS+NAA 0.15 mg/L+6-BA 0.3 mg/L培养基激素配比适宜,能诱导较多的气生鳞茎;经4℃低温贮藏60 d后,大蒜花苞在MS+NAA 0.1 mg/L+ABA 1.0 mg/L激素配比培养基上气生鳞茎诱导率可达95.0%。大蒜气生鳞茎萌发受到其个体大小的影响,较小的气生鳞茎能迅速且萌发率高,较大的气生鳞茎则萌发率低。气生鳞茎在MS+NAA 0.05 mg/L+6-BA 0.1 mg/L培养基激素配比较适宜,能诱导较多的气生鳞茎萌发。(3)扫描电镜检测结果表明,未进行茎尖培养前,成县迟蒜被马铃薯Y病毒属、青葱潜隐病毒属、大蒜潜隐病毒属、麝香石竹潜隐病毒属的病毒侵染;鲁蒜王Ⅱ号被麝香石竹潜隐病毒属、马铃薯Y病毒属、一些长线病毒侵染;在经过茎尖脱病毒后,在两个品种中只检测到少量麝香石竹潜隐病毒属和马铃薯Y病毒属。表明在本研究范围内,大蒜茎尖培养配合热处理可对大蒜病毒进行有效脱除,脱毒率达到60%以上。更多还原

【Abstract】 Garlic (Allium sativum L.) is an important vegetable crops, the production of asexual reproduction mainly by the bulb. Garlic in the long process of asexual reproduction , accumulation of susceptible viruses and widespread, resulting in degradation of caste, quality and yield. Garlic apical meristem removal combined with heat treatment is an effective method of garlic virus, the virus can be effectively removed most of the substantial increase in the quality and yield of garlic. However, difficult the garlic shoot tip culture propagation coefficient low, virus identification difficulties, virus-free seed production costs high, a virus-free garlic seed cultivation and production of application bottlenecks.This research shoot apical meristem culture of garlic with heat treatment approach removal of viruses,and to establish the garlic shoot tips virus-free culture system. And substitute with fresh or low temperature storage of garlic sprouts as explants to study the storage time and culture of plant growth hormones induce the formation of inflorescence axis of bulbils, and the size of bulbils, temperature and plant hormone on aerial germination rate of the bulbs. Detected by scanning electron microscopy before and after tissue culture of garlic removal virus contained in the virus situation.(1) Two species of garlic bulbs by the line combination of heat treatment and detoxification of tip growing point.The results showed that MS +0.1 mg / L NAA +0.5 mg/L6-BA garlic shoot tips on medium had a higher germination rate and in vitro bulb formation rate, but also a strong growth potential, germination rate and in vitro bulb formation rate up to 97.4% and 87.2%; 30d at 37℃heat treatment after the bulb of garlic shoot tips have a higher formation rate; cut the size of 0.3 ~ 0.5mm tip, the tip have a higher survival rate, bulb formation rate and growth potential.(2)The inflorescence of garlic can be used as an explant in vitro to formed bulblets. After low temperature storage, formation rate of bulblets is relatively low. Formation of bulblets germinated by the impact of bulblets size. By MS as medium,with no storage and storage30,60,90and120d, respectively, the inflorescences of garlic were used as explants for tissue culture.The results indicated that the formation rate of bulblets could reach 95.0% under the interactions treatments of MS+NAA 0.1 mg/L+ABA 1.0 mg/L,after 60 d storage at 4℃. Small bulblets can germinate rapidly and theirs shooting rate is high reached 77.9%,but big bulblets come to the opposite.The MS+ NAA 0.05 mg/L+6-BA 0.1 mg/L medium is appropriate proportion,which can induce more bulblet shooting(3)In order to provide theroretical guidance for screening no virus plants and application, electron microscope technology is used to detect the virus of the Late garlic from Gansu and King of garlic Lu from Shandong.The results showed in the Late garlic from Gansu it had Potyvirus,Shallot latent virus(ShLY), Garlic common latent virus and Carlavirus;in King of garlic Lu from Shandong garlic it had Shallot latent virus(ShLY), Potyvirus and many different long length virus. That within the scope of this study, treatment with garlic shoot tip culture of garlic can be effective virus removal, virus-free rate of 60% or more.更多还原