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ABSTRACT: Tea is the most consumed beverage worldwide after water. Yet very little is known about the genetics of tea in comparison with other crop species. Here we have taken advantage of the polymorphic nature of microsatellite DNA to investigate the mode of chloroplast inheritance in tea, Camellia sinensis (L.) O. Kuntze. This is important for the correct interpretation of phylogeny and introgression data as well as assessing the suitability of chloroplast transformation as a means for transgene containment in tea.
The study was based on six Japanese tea cultivars, namely Aj2, CK23, Hatsumomiji, Nka05, Yamanoibuki and Kanayamidori used to generate four informative families. The parental pairs in the crosses differed at a single chlroroplast locus with respect to an imperfect microsatellite repeat of 16 nucleotide bases. In agreement with earlier cytological studies, all 61 progeny displayed a cpDNA profile that was consistent with the maternal inheritance of chloroplasts in tea.
The data generated here provide the first molecular evidence of the plastid inheritance in tea. However, we suggest that additional families and polymorphic markers be screened for increasing the confidence in the observed maternal inheritance of chloroplasts in this important crop species.
Journal of the Science of Food and Agriculture 07/2011; 91(14):2660-3. · 1.44 Impact Factor
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ABSTRACT: We have developed PCR-based DNA markers with the objective of authenticating, and discouraging the fraudulent commercialization of, the forty-four most cultivated Japanese green teas. For this purpose we have generated amplicon length polymorphisms (ALPs) by targeting microsatellite loci using nucleotide information from other species and indels located in the non-coding regions of phenylalanine ammonia-lyase (PAL) and dihydroflavonol 4-reductase (DFR) genes in tea. One useful indel was detected in the unique intron of PAL and three others in introns 2, 3 and 5 of the DFR gene. All four nuclear microsatellite loci were polymorphic showing three to four alleles per locus, while only one out of the four chloroplast loci was variable with two distinct haplotypes. PCR primers were designed to amplify small but variable PCR fragments in such a way that they were applicable to low amounts of highly degraded DNA. The markers devised here were sufficient to differentiate all the teas, although a combination of several loci was necessary for most of them. This single-step PCR-based methodology is more effective than the PCR-RFLP technique commonly used for identifying food materials and will be useful for the certification of commercial Japanese green teas. Copyright © 2004 Society of Chemical Industry
Journal of the Science of Food and Agriculture 05/2004; 84(8):895 - 902. · 1.44 Impact Factor
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ABSTRACT: In Japan, tea is generally sold blended, though 90% of the total production is clonal. Due to the increasingly strict consumer need and taste, however, more and more Japanese green teas are being sold under their particular cultivar name. Moreover, tea made from Yabukita, a much appreciated cultivar originally developed in Japan, has recently been produced and imported from a neighboring country. This paper describes a simple and inexpensive methodology capable of identifying fresh and processed Japanese green teas to discourage its fraudulent commercialization. The study was based on 46 main tea cultivars, and polymorphism detected through STS-RFLP analysis of the coding and noncoding DNA regions of three genes, namely phenylalanine ammonia-lyase, chalcone synthase, and dihydroflavonol 4-reductase, for which nucleotide information was available. All 46 tea cultivars analyzed could be easily distinguished using a combination of codominant DNA markers. Yabukita displayed a unique profile when PAL intron was digested with DdeI, thus allowing its rapid authentication at low cost.
Journal of Agricultural and Food Chemistry 04/2003; 51(7):1765-70. · 2.82 Impact Factor
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ABSTRACT: The genetic diversity of tea, Camellia sinensis (L.) O. Kuntze, including the two main cultivated sinensis and assamica varieties, was investigated based on PCR-RFLP analysis of PAL, CHS2 and DFR, three key genes involved in catechin and tannin synthesis and directly responsible for tea taste and quality. Polymorphisms were of two types: amplicon length polymorphism (ALP) due to the presence of indels in two introns of PAL and DFR, and point mutations detected after restriction of amplified fragments with appropriate enzymes. A progeny test showed that all markers segregated in a Mendelian fashion and that polymorphisms were exclusively co-dominant. CHS2, which belongs to a multi-gene family, allowed for greater variation than the single-copy PAL gene. Based on Nei's gene diversity index, var. sinensis was revealed to be more variable than var. assamica, and that a higher proportion of overall diversity resided within varieties as compared to between varieties. Even though no specific DNA profile was found for either tea varieties following any single PCR-RFLP analysis, a factorial correspondence analysis carried out on all genotypes and markers separated the tea samples into two distinct groups according to their varietal status. This reflects the large difference between var. sinensis and var. assamica in their polyphenolic profiles. The STS-based markers developed in this study will be very useful in future mapping, population genetics and fingerprinting studies of this important crop species and other Camellia species, as the primers have also proven successful in the three other subgenera of this genus.
Theoretical and Applied Genetics 03/2003; 106(3):375-83. · 3.30 Impact Factor
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ABSTRACT: The advantage of the cross transferability of heterologous chloroplast and nuclear microsatellite primers was taken to detect polymorphism among 24 tea (Camellia sinensis (L.) O. Kuntze) genotypes, including both the assamica and the sinensis varieties. Primer information was obtained from the closely related Camellia japonica species for four nuclear microsatellites, and from Nicotiana tabaccum for seven universal chloroplast microsatellites. All of the nuclear microsatellite loci tested generated an expected DNA fragment in tea, revealing between three and five alleles per locus. Four out of the seven chloroplast microsatellites primers amplified positively, and of these only one was polymorphic with three alleles, which is in agreement with the conserved nature of chloroplast microsatellites at the intraspecific level. A factorial correspondence analysis carried out on all genotypes and nuclear microsatellite alleles separated the assamica and sinensis genotypes into two groups, thus demonstrating the value of these markers in establishing the genetic relationship between tea varieties. Genetic diversity measured with nuclear microsatellites was higher than that measured with other types of molecular markers, offering prospects for their use in fingerprinting, mapping, and population genetic studies, whereas polymorphisms detected at a cpSSR locus will allow the determination of plastid inheritance in the species.
Genome 01/2003; 45(6):1041-8. · 1.65 Impact Factor
