Functional study of CHS gene family members in citrus revealed a novel CHS gene affecting the production of flavonoids.

BACKGROUND: Citrus flavonoids are considered as the important secondary metabolites because of their biological and pharmacological activities. Chalcone synthase (CHS) is a key enzyme that catalyses the first committed step in the flavonoid biosynthetic pathway. CHS genes have been isolated and characterized in many plants. Previous studies indicated that CHS is a gene superfamily. In citrus, the number of CHS members and their contribution to the production of flavonoids remains a mystery. In our previous study, the copies of CitCHS2 gene were found in different citrus species and the sequences are highly conserved, but the flavonoid content varied significantly among those species. RESULTS: From seventy-seven CHS and CHS-like gene sequences, ten CHS members were selected as candidates according to the features of their sequences. Among these candidates, expression was detected from only three genes. A predicted CHS sequence was identified as a novel CHS gene. The structure analysis showed that the gene structure of this novel CHS is very similar to other CHS genes. All three CHS genes were highly conserved and had a basic structure that included one intron and two exons, although they had different expression patterns in different tissues and developmental stages. These genes also presented different sensitivities to methyl jasmonate (MeJA) treatment. In transgenic plants, the expression of CHS genes was significantly correlated with the production of flavonoids. The three CHS genes contributed differently to the production of flavonoids. CONCLUSION: Our study indicated that CitCHS is a gene superfamily including at least three functional members. The expression levels of the CHS genes are highly correlated to the biosynthesis of flavonoids. The CHS enzyme is dynamically produced from several CHS genes, and the production of total flavonoids is regulated by the overall expression of CHS family genes.

A 5'-proximal region of the Citrus tristeza virus genome encoding two leader proteases is involved in virus superinfection exclusion.

Superinfection exclusion (SIE), a phenomenon in which a primary virus infection prevents a secondary infection with the same or closely related virus, has been observed with various viruses. Earlier we demonstrated that SIE by Citrus tristeza virus (CTV) requires viral p33 protein. In this work we show that p33 alone is not sufficient for virus exclusion. To define the additional viral components that are involved in this phenomenon, we engineered a hybrid virus in which a 5'-proximal region in the genome of the T36 isolate containing coding sequences for the two leader proteases L1 and L2 has been substituted with a corresponding region from the genome of a heterologous T68-1 isolate. Sequential inoculation of plants pre-infected with the CTV L1L2T68 hybrid with T36 CTV resulted in superinfection with the challenge virus, which indicated that the substitution of the L1-L2 coding region affected SIE ability of the virus.

Marker-Assisted Selection of Tomato Genotypes with the I-2 Gene for Resistance to Fusarium oxysporum f. sp. lycopersici Race 2.

Fusarium vascular wilt, caused by Fusarium oxysporum f. sp. lycopersici, affects tomatoes worldwide. Development of resistant varieties of tomato would constitute an economically and environmentally sound approach for the management of this disease. Resistance genes to F. oxysporum f. sp. lycopersici race 1 (I-1 gene) and race 2 (I-2 gene) were mapped to chromosome 11. The I-2 gene cluster includes one functional copy and six nonfunctional homologs of the I-2 gene. This report describes the design of primers based on the functional gene copy and the development of a multiplex polymerase chain reaction (PCR)-based method that has the ability to differentiate I-2 genotypes from genotypes without the I-2 gene. In these trials, 39 of the 40 genotypes tested with known reactions to race 2 gave the expected results. The only exception was the cultivar Plum Crimson carrying the I-3 gene for resistance, which confers resistance to F. oxysporum f. sp. lycopersici races 1, 2, and 3. This method was validated in three countries and by bioassays.