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 หัวเรื่อง:ไม่มีชื่อไทย (ชื่ออังกฤษ : Development of Microsatellite Markers for Vanda Orchid) ผู้เขียน: ปรารถนา เผือกวิไล,  นายประดิษฐ์ พงศ์ทองคำ, ศาสตราจารย์เกียรติคุณ , สุรินทร์ ปิยะโชคณากุล สื่อสิ่งพิมพ์:pdf AbstractThe aim of this research was to develop microsatellite (SSR) markers for the Vanda orchid from the enriched library using dinucleotide repeats [(CA)15 and (GA)15] and trinucleotide repeats [(ACC)10 and (CCT)10] as probes. Positive clones were selected using dot blot hybridization. The results showed that 82.45% of dinucleotide-enriched libraries but only 9.91% of trinucleotide-enriched libraries gave positive signals. After sequencing, 83.12% of the positive clones contained microsatellite repeats. The four most frequently found sequences were the compound repeats of (GA)n(GT)n (45.19%), (GA)n (22.59%), (CA)n (15.93%) and (CCT)n (9.26%). Fifty-six pairs of primers were designed and nine primer pairs could amplify the DNA giving the expected PCR product with polymorphism. There was a range from 3 to 9 alleles per locus and the expected heterozygosity (He) range was 0.3150-0.7438. Based on the nine loci of these microsatellite markers, the probability of identity (PI) of any two Vanda and related orchid cultivars having the same genotype was approximately 1 in 1,000,000. Therefore, these markers could be used for identification of the Vanda orchid samples studied. After analyzing the genetic relationships of 33 Vanda and related orchid cultivars using NTSYS-pc 2.1m, the result indicated that the Vanda and related orchid cultivars could be divided into four groups. The first three groups were strap-leaved Vanda, while the fourth one was terete Vanda, which was clearly clustered separately from the other Vanda groups. This study showed the isolation efficiency of the enrichment procedure, the abundance of microsatellites in Vanda orchids and their potential use for the individual identification of Vanda and related orchid samples. |
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หัวเรื่อง:ไม่มีชื่อไทย (ชื่ออังกฤษ : Sequence Analysis of Ethylene Response Sensor Gene Isolated from Vanda Miss Joaquim Flower ) ผู้เขียน:ดร.นพมาศ โลกคำลือ, ผู้ช่วยศาสตราจารย์ , พัฒนา ศรีฟ้า ฮุนเนอร์ สื่อสิ่งพิมพ์:pdf AbstractA specific ethylene receptor protein is a major key for ethylene perception into plant tissues. Thus, the Ethylene Response Sensor (ERS) gene was isolated from the ethylene-sensitive flowers of Vanda Miss Joaquim and its deduced ethylene response sensor (ERS) was aligned with three other reported orchid ERS genes to identify the specific divergence in orchid ethylene receptor proteins. The results from multiple alignment showed that the amino acid sequence of the Vanda ERS was 95% identical to Phalaenopsis ERS (the ethylene-sensitive species), but shared 90–91% identity with the ERS isolated from two ethylene-insensitive orchid species, Dendrobium hybrid Khao Sanan and Oncidium Gower Ramsey. The most conserved regions were located at the N-terminus of the polypeptide being three transmembrane hydrophobic regions; however, the most variable regions, particularly for the Dendrobium ERS and Oncidium ERS, were located in the ATP-binding site of the histidine kinase domain and the amino acid sequence of the downstream histidine kinase domain at the C-terminus. The findings suggest the sequence similarity in the ATP-binding site can be used to distinguish the closely related ERS proteins. In considering the level of ERS gene expression, there was an abundance of ERS transcript accumulation in fully opened Vanda Miss Joaquim flowers (particularly in the column and the lip followed by the perianth tissue) compared with root and leaf tissues. |
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