Application of HAT-RAPD Technique in Identifying Natural Hybrids of Roscoea (Zingiberaceae)

Natural hybridization is very common in plants, and plays an important role in plant evolution. Besides the traditional methods including morphological analysis and hand crossing, molecular evidence is needed for studying natural hybridization.In order to analyze natural hybridization in Roscoea,HAT-RAPD technique was used toidentify putative hybrids from parental species by principal co ordinate analysis and hybrid index. The results indicated that the bands amplified by HAT-RAPD technique were more stable and reliable than that of RAPD. The result of principal co ordinate analysis and hybrid index showed that intermediate individuals were the hybrids of R.humeana and R.cautleoides, and showed closer relationships to R.humeana. These results suggested that HAT-RAPD could be used to study natural hybridization. As it is simple and easy to manipulate, HAT RAPD may prove to be a very effective technique in hybrid identification in the studies of plant evolution.     

HAT-RAPD技术在姜科象牙参属植物自然杂交个体鉴定中的应用

自然杂交在植物中广泛存在,对植物进化起着重要的作用.研究自然杂交现象,除了传粉、形态等传统手段,还需要分子水平的证据.本研究利用HAT-RAPD技术,对姜科象牙参属植物的自然杂交现象进行分析,运用主坐标分析和杂交指数分析对疑似杂交个体进行鉴定,探讨HAT-RAPD技术在分析和鉴定杂交个体方面的可行性.结果显示,HAT-RAPD方法比传统RAPD扩增的条带更稳定、清楚、容易统计,同时主坐标分析和杂交指数分析的结果表明,形态介于大花象牙参(Roscoea humeana)和早花象牙参(R.cautleoides)的个体是两者的杂交后代,且与大花象牙参的亲缘关系更近.以上的结果表明HAT-RAPD技术可以用于自然杂交的分析,并且由于其简单、易操作等特点,将成为一种非常适合分析和鉴定杂交个体的分子手段.     

A reconsideration of relationships among Japanese Trillium species based on karyology and AFLP data

To reexamine the relationships among the Japanese Trillium species that form a polyploid series, we performed principal-coordinates analysis (PCOA) based on proposed karyotypic compositions and on amplified fragment length polymorphism (AFLP) analysis. A hexaploid species, T. smallii, whose karyotypic composition had been hypothesized as K₂K₂SSUU, with hybridization between tetraploid T. apetalon (SSUU) and a presumed K₂K₂ diploid species, showed a genotype corresponding to T. yezoense (K₁SU). Accordingly, T. smallii appears to be an allopolyploid of T. yezoense, with the karyotypic composition K₁K₁SSUU. Trillium channellii, a recently described tetraploid species whose origin and genealogical position remains unclear, showed a genotype corresponding to K₁K₁K₂T. We conclude that T. channellii may be derived from hybridization of T. camschatcense (K₁K₁) as the seed parent and T. tschonoskii (K₂K₂TT) or hexaploid T. hagae (K₁K₁K₂K₂TT) as the pollen parent.     

Analysis of intraspecific and interspecific variation in the genus Alternaria by the use of RAPD-PCR

The RAPD-PCR profiles of 13 phytopathogenic Alternaria species and two closely related outgroups were examined using six different primers. Each species produced a distinct pattern of DNA fragments which were used as a measure of the degree of relatedness between species. A. brassicae isolates of diverse origin showed high levels of similarity but little similarity was noted between other species. The closest interspecific genetic distances were recorded between A. citri, A. alternata and A. longipes. The outgroup genera Embellisia and Stemphylium, which are recognised as distinct, could not be clearly separated using RAPD banding criteria, suggesting a high level of genetic diversity amongst these groups of fungi.     

Discrimination of cassava-associated Bemisia tabaci in Africa from polyphagous populations, by PCR–RFLP of the internal transcribed spacer regions of ribosomal DNA

Restriction fragment length polymorphism (RFLP) analysis of the ribosomal DNA internal transcribed spacer regions of Bemisia tabaci was used to distinguish cassava‐associated populations from other host‐associated populations. Endonuclease restriction profile analysis indicated that cassava‐associated populations from Africa represent a distinct group, with a significant level of separation into subgroups that were not linked to geographical origin. Analysis of molecular variance (amova ) revealed that a high proportion of the total genetic variation (47%) was attributable to among‐population differences within the host‐associated groups. Principal coordinate analysis supported the differentiation between the cassava and the non‐cassava group, a result which was in agreement with the cluster analysis of the restriction fragment profile. Internal transcribed spacer RFLP markers, especially SmaI, identified in this study can be used to monitor the spread of B. tabaci biotypes, especially of the more virulent biotype B that has so far not been reported in the cassava‐growing belt of Africa.