茶菊病毒检测与脱除技术研究

为了检测茶用菊品种病毒感染情况,并研究产业面积较大的福白菊的脱毒技术。本试验采用巢式PCR方法对24个茶用菊品种进行菊花常见8种(类)病毒的检测,比较了5种不同浓度6-苄基腺嘌呤(6-BA)和萘乙酸(NAA)组合的MS培养基对福白菊茎尖的诱导效果,并用筛选出的激素配比结合4℃低温和利巴韦林处理对福白菊进行脱毒;采用SRAP分子标记技术检测福白菊的脱毒株与非脱毒株,并检测植株生长指标、花朵产量以及花朵总黄酮、绿原酸、木樨草苷、异绿原酸A含量,比较脱毒苗与非脱毒苗的差异。结果表明,24个茶用菊品种中感染较为普遍的有两种病毒(CVB、TAV)和两种类病毒(CSVd和CChMVd),其中,感染率最高的为CSVd,高达100%,其次是TAV(91.67%)、CVB(87.50%),而CChMVd的感染率为66.67%。福白菊单株则感染了CVB和CSVd,福白菊茎尖在1 mg·L^-1 6-BA和0.1 mg·L^-1 NAA的MS培养基中生长健壮且出愈率最高。脱毒后的福白菊保持了遗传稳定性,采用脱除病毒的福白菊母本扦插繁育的种苗生产较源自非脱毒种源生产的花朵产量、品质等指标都有显著提高。综上所述,茶用菊品种普遍感染各种病毒,利用茎尖培养可以对福白菊为代表的茶用菊脱毒,脱毒之后福白菊花朵产量和品质均明显提高。本研究结果为茶用菊的脱毒和种源复壮提供了有效方法。

Research of Virus Detection and Elimination of Tea Chrysanthemum

To detect the virus infection of different tea chrysanthemum varieties, widely planted tea chrysanthemum Fubaiju was selected as material to optimize the virus elimination technology, and nested PCR was used to detect eight virus or viroids in 24 varieties of tea chrysanthemum; five kinds of MS media containing ribavirin with different concentrations of 6-benzylaminopurine(6-BA) and 1-naphthaleneacetic acid(NAA) were used to optimize the culture conditions of Fubaiju shoot under 4℃, then the induction effects were examined; SRAP molecular marker technology was used to detect the genetic differences between the virus-free and non-virus-free seedlings of Fubaiju, further the flower yield and the content of total flavonoids, chlorogenic acid, luteoloside and isochlorogenic acid A were compared between this two strains. The results show that two kinds of virus (CVB, TAV) and two kinds of viroids (CSVd and CChMVd) were widespread in 24 tea chrysanthemum varieties. The infection rates of CSVd, TAV, CVB and CChMVd were 100%, 91.67%, 87.50% and 66.7%, respectively. The Fubaiju plants were infected by CVB and CSVd. The shoot of Fubaiju grew healthily and produced more calluses in MS medium containing 1 mg·L^-1 6-BA and 0.1 mg·L^-1 NAA. The virus-free Fubaiju plants maintained its genetic stability, and the yields and qualities of flowers were significantly improved compared with those of the non-virus-free ones. In brief, the infection of virus widely spread in tea chrysanthemum varieties; the virus and viroid in Fubaiju can be effectively eliminated using shoot culture technology with significant improvement of their flower yield and quality. The results provide an effective method for virus elimination and rejuvenation of tea chrysanthemum.