Sota Koeda

Dramatic changes in leaf development of the native Capsicum chinense from the Seychelles at temperatures below 24°C

When a pepper cultivar (Capsicum chinense cv. Seychelles-2, Sy-2) native to the Seychelles was grown in Japan, all seedlings showed seasonal developmental abnormalities such as development of abnormally shaped leaves. Other pepper cultivars grew well in all seasons while the growth of cv. Sy-2 was stunted. In this study, we first examined the effects of various changes in temperature and photoperiod on the cv. Sy-2 phenotype. The results showed that temperatures lower than 24°C led to the formation of abnormal leaves. Second, morphological and anatomical analyses of cotyledons and true leaves developed at 28 and 20°C were conducted. The narrower and thicker cotyledons developed at 20°C had fewer palisade cells in the leaf-length direction, and more cells in the leaf-thickness direction. True leaves developed at 20°C were irregularly shaped, thicker and had smaller leaf area. In addition, true leaves developed at 20°C had fewer palisade cells in the leaf-length and leaf-width directions and had more cells in the leaf-thickness direction. Furthermore, abnormal periclinal cell divisions in the mesophyll and/or epidermal cell layers were observed during leaf blade development at 20°C. These results suggest that the observed changes in cell proliferation and abnormal periclinal cell divisions were related, at least in part, to abnormal leaf development of cv. Sy-2 at temperatures below 24°C.

Multiple loss-of-function putative aminotransferase alleles contribute to low pungency and capsinoid biosynthesis in Capsicum chinense

Capsicum chinense is a domesticated hot pepper species in the Capsicum genus that originated in the Amazon and is consumed in USA, the Caribbean and South America. Although a characteristic of this species is high pungency, some non-pungent or low-pungent strains, called “Aji Dulce” (sweet pepper in Spanish), exist in the Caribbean region. In the present study, low-pungent C. chinense accessions were analyzed in order to elucidate the genetic mechanisms responsible for low pungency. All low-pungent C. chinense accessions in this study carried non-functional alleles of putative aminotransferase (pAMT), which catalyzes the formation of vanillylamine from vanillin in the capsaicinoid biosynthetic pathway. These low-pungent accessions produced capsinoids, low-pungent capsaicinoid analogs. The pamt mutation in each strain was characterized using allele-specific markers, and one novel pamt allele (pamt7) was identified. The pamt7 had a new hAT family transposon insertion in the second exon region, which caused the loss of pAMT expression. pamt7 is apparently an ancestral allele for pamt6 because the 7-bp insertion in pamt6 can be regarded as a footprint of the transposon. A phylogenetic analysis of pamt alleles was performed to examine their relationships. Combined with previously reported pamt alleles, the Tcc family transposon insertion and its excision were involved in the generation of various pamt alleles in C. chinense. A phylogenetic analysis of pamt alleles showed that at least five occurred within C. chinense after speciation of the Capsicum genus. In conclusion, the results of the present study identified pamt as the main and most frequent gene controlling low pungency in C. chinense. Allelic variations in loss-of function pamt and their wide distribution demonstrated the potential of C. chinense bioresources for genetic improvements to pungency and metabolic profiles in hot pepper breeding programs.

Defense response of a pepper cultivar cv. Sy-2 is induced at temperatures below 24°C.

Temperature is one of the most important environmental factors that influence plant growth and development. Recent studies imply that plants show various responses to non-extreme ambient temperatures. Previously, we have found that a pepper cultivar cv. Sy-2 (Capsicum chinense) shows developmental defects at temperatures below 24°C. In this study, to gain new insights into the temperature sensitivity of cv. Sy-2, temperature-sensitive genes were screened using microarray techniques. At restrictive temperature of 20°C, almost one-fourth of the 411 up-regulated genes were defense related or predicted to be defense related. Further expression analyses of several defense-related genes showed that defense-related genes in cv. Sy-2 were constitutively expressed at temperatures below 24°C. Moreover, accumulation of high level of salicylic acid (SA) in cv. Sy-2 grown at 20°C suggests that the defense response is activated in the absence of pathogens. To confirm that the defense response is induced in cv. Sy-2 below 24°C, we evaluated the resistance to biotrophic bacterial pathogen Xanthomonas campestris pv. vesicatoria and necrotrophic fungal pathogen Cercospora capsici. Cv. Sy-2 showed enhanced resistance to X. campestris pv. vesicatoria, but not to C. capsici.