Effect of milk types on the attributes of a glutinous rice wine-fermented yogurt-like product.

This study was conducted to analyze the effect of milk types on the attributes of the glutinous rice wine-fermented yogurt-like product named Kouwan Lao (KWL). Four types of raw milks were used in this study, including high temperature, long time (HTLT: H milk), HTLT milk supplemented with 3% skim milk powder (S milk), pasteurized milk (P milk), and ultra-high temperature milk (U milk). Microbiological compositions of the fermented glutinous rice and KWL at different stages were analyzed using PCR-denaturing gradient gel electrophoresis and gene sequencing based on 16S rRNA and 26S rRNA. The physicochemical properties of KWL samples were determined, and textural properties of those were analyzed using a texture analyzer (Jiawei Innovation and Technology Co. Ltd., Zhejiang Province, China). The microstructure of KWL samples was observed using scanning electron microscopy. The results showed that the milk types had significant influences on the bacterial composition of KWL. In the curdling process, the predominant bacteria of H, S, P, and U KWL samples were Lactobacillus brevis, Janthinobacterium sp., Lactobacillus casei, and Streptococcus agalactiae, respectively. In the ripening process, the main strains in H KWL were Enterococcus faecium and Pediococcus pentosaceus. Lactobacillus casei and Lactobacillus paracasei were the dominant bacteria of U KWL. Lactobacillus casei was the main strain of P KWL, and no bacteria were detected in S KWL. Saccharomyces cerevisiae was the dominant fungus of KWL, and no significant effect of milk types on fungal composition of KWL was found. The results of physicochemical properties showed that significant differences in protein contents were found in the KWL samples, and P KWL had the highest protein content. The fat content of U KWL was significantly higher than that of samples from the other 3 groups. The U KWL and P KWL showed lower moisture than that of the other 2 KWL samples. In addition, no significant difference in pH value was found in all samples. The results of texture analysis and microstructure showed that compared with other 3 types of KWL samples, the related mass parameters of U KWL were more advantageous and improved significantly with the increase of the heat treatment temperature of raw milk and the addition of skim milk powder. Our findings revealed the effects of milk types on microbial composition, physicochemical properties, textural properties, and microstructure of KWL, and provided a basic theory for the optimization and industrial production of KWL.

Genetic mapping and development of molecular markers for a candidate gene locus controlling rind color in watermelon.

KEY MESSAGE: ClCG08G017810 (ClCGMenG) encoding a 2-phytyl-1,4-beta-naphthoquinone methyltransferase protein is associated with formation of dark green versus light green rind color in watermelon. Rind color is an important agronomic trait in watermelon [Citrullus lanatus (Thunb.) Matsum. and Nakai], but the underlying molecular mechanism for this trait is not fully known. In the present study, we identified a single locus on chromosome 8 accounting for watermelon rind color (dark green vs. light green). Genetic analysis of F1, F2, and BC1 populations derived from two parental lines (9904 with dark green rind and Handel with light green rind) revealed that the watermelon rind color (dark green vs. light green) is controlled by a single locus, and dark green is dominant to light green rind. Initial mapping revealed a region of interest spanning 2.07 Mb on chromosome 8. Genetic mapping with CAPS and SNP markers narrowed down the candidate region to 31.4 kb. Gene annotation of the corresponding region in the reference genome revealed the ClCG08G017810 gene sequence encoding the 2-phytyl-1,4-beta-naphthoquinone methyltransferase protein. The sequence alignment of the candidate gene with the two parental lines suggested a nonsynonymous SNP mutation in the coding region of ClCG08G017810, converting an arginine (R) to glycine (G). The SNP might be associated with rind color of 103 watermelon germplasm lines investigated in this study. The qRT-PCR analysis revealed higher expression of ClCG08G017810 in dark green rind than in light green rind. Therefore, ClCG08G017810 is a candidate gene associated with watermelon rind color. The present study facilitates marker-assisted selection useful for the development of cultivars with desirable rind color.

Genetic mapping reveals a candidate gene (ClFS1) for fruit shape in watermelon (Citrullus lanatus L.).

KEY MESSAGE: A 159 bp deletion in ClFS1 gene encoding IQD protein is responsible for fruit shape in watermelon. Watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai] is known for its rich diversity in fruit size and shape. Fruit shape has been one of the major objectives of watermelon breeding. However, the candidate genes and the underlying genetic mechanism for such an important trait in watermelon are unknown. In this study, we identified a locus on chromosome 3 of watermelon genome controlling fruit shape. Segregation analysis in F2 and BC1 populations derived from a cross between two inbred lines "Duan125" (elongate fruit) and "Zhengzhouzigua" (spherical fruit) suggests that fruit shape of watermelon is controlled by a single locus and elongate fruit (OO) is incompletely dominant to spherical fruit (oo) with the heterozygote (Oo) being oval fruit. GWAS profiles among 315 accessions identified a major locus designated on watermelon chromosome 3, which was confirmed by BSA-seq mapping in the F2 population. The candidate gene was mapped to a region 46 kb on chromosome 3. There were only four genes present in the corresponding region in the reference genome. Four candidate genes were sequenced in this region, revealing that the CDS of Cla011257 had a 159 bp deletion which resulted in the omission of 53 amino acids in elongate watermelon. An indel marker was derived from the 159 bp deletion to test the F2 population and 105 watermelon accessions. The results showed that Cla011257 cosegregated with watermelon fruit shape. In addition, the Cla011257 expression was the highest at ovary formation stage. The predicted protein of the Cla011257 gene fitted in IQD protein family which was reported to have association with cell arrays and Ca2+-CaM signaling modules. Clear understanding of the genes facilitating the fruit shape along with marker association selection will be an effective way to develop new cultivars.

Genetic mapping reveals a marker for yellow skin in watermelon (Citrullus lanatus L.).

As a diverse species, watermelon [Citrullus lanatus (Thunb.) Matsum. &Nakai var. lanatus] has different kinds of fruit sizes, shapes, flesh colors and skin colors. Skin color is among the major objectives for breeding. Yellow skin is an important trait in watermelon, but the underlying genetic mechanism is unknown. In this study, we identified a locus for yellow skin through BSA-seq and GWAS. A segregation analysis in F2 and BC1 populations derived from a cross of two inbred lines '94E1'(yellow skin) and 'Qingfeng'(green skin) suggested that skin color is a qualitative trait. BSA-seq mapping confirmed the locus in the F2 population, which was detected on chromosome 4 by GWAS among 330 varieties. Several major markers, namely, 15 CAPS markers, 6 SSR markers and 2 SNP markers, were designed to delimit the region to 59.8 kb region on chromosome 4. Utilizing the two populations consisting of 10 yellow and 10 green skin watermelons, we found a tightly linked functional SNP marker for the yellow skin phenotype. The application of this marker as a selection tool in breeding programs will help to improve the breeder's ability to make selections at early stages of growth, thus accelerating the breeding program.