Identification, Characterization and Expression Analysis of Anthocyanin Biosynthesis-related bHLH Genes in Blueberry (Vaccinium corymbosum L.)

Basic helix-loop-helix proteins (bHLHs) play very important roles in the anthocyanin biosynthesis of many plant species. However, the reports on blueberry anthocyanin biosynthesis-related bHLHs were very limited. In this study, six anthocyanin biosynthesis-related bHLHs were identified from blueberry genome data through homologous protein sequence alignment. Among these blueberry bHLHs, VcAN1, VcbHLH42-1, VcbHLH42-2 and VcbHLH42-3 were clustered into one group, while VcbHLH1-1 and VcbHLH1-2 were clustered into the other group. All these bHLHs were of the bHLH-MYC_N domain, had DNA binding sites and reported conserved amino acids in the bHLH domain, indicating that they were all G-box binding proteins. Protein subcellular location prediction result revealed that all these bHLHs were nucleus-located. Gene structure analysis showed that VcAN1 gDNA contained eight introns, while all the others contained seven introns. Many light-, phytohormone-, stress- and plant growth and development-related cis-acting elements and transcription factor binding sites (TFBSs) were identified in their promoters, but the types and numbers of cis-elements and TFBSs varied greatly between the two bHLH groups. Quantitative real-time PCR results showed that VcAN1 expressed highly in old leaf, stem and blue fruit, and its expression increased as the blueberry fruit ripened. Its expression in purple podetium and old leaf was respectively significantly higher than in green podetium and young leaf, indicating that VcAN1 plays roles in anthocyanin biosynthesis regulation not only in fruit but also in podetium and leaf. VcbHLH1-1 expressed the highest in young leaf and stem, and the lowest in green fruit. The expression of VcbHLH1-1 also increased as the fruit ripened, and its expression in blue fruit was significantly higher than in green fruit. VcbHLH1-2 showed high expression in stem but low expression in fruit, especially in red fruit. Our study indicated that the anthocyanin biosynthesis regulatory functions of these bHLHs showed certain spatiotemporal specificity. Additionally, VcAN1 might be a key gene controlling the anthocyanin biosynthesis in blueberry, whose function is worth exploring further for its potential applications in plant high anthocyanin breeding.

Pemetaan Atribut Sensori Kopi Kobura Berdasarkan Perbedaan Cara Panen dan Pengolahan di Tingkat Petani

<p><em>In general, Kobura coffee farmers in South Ogan Komering Ulu (OKU) harvest coffee with the criteria of red fruit and a mixture of red and green, while the processing method is a dry process. However, the sensory attributes of these activities are not yet known. This study aims to map the sensory qualities of Kobura coffee at the farmer level based on differences in harvesting and processing methods. The research was carried out in Banding Agung District, South OKU Regency, and the Coffee and Cocoa Bioindustry Laboratory, Balittri, Sukabumi, from March to December 2019. The design used was a completely randomized design with 3 treatment factors and two replications. The first factor was three clones of Kobura coffee (Kobura 1, 2, 3), the second factor was two harvesting methods (red and mix of red + green fruit), and the third factor is two processing methods (dry and wet). The variable observed were the sensory attributes analyzed by multivariate analysis of variance, principal component analysis, cluster analysis, and path analysis. The results showed that the harvesting and processing methods of Kobura coffee at the farmer level in South OKU did not significantly affect the sensory. Furthermore, each cluster's harvesting and processing methods were evenly distributed because the proportion between red and green fruit was not clearly identified in the mixed fruit harvest. Superior genetic and optimal altitude factors make Kobura coffee categorized into Fine Robusta with a final score of 81.13-85.75. The final score is directly and dominantly influenced by mouthfeel (body), flavor, overall, and fragrance, respectively.</em></p>

Comprehensive Analysis of the SBP Family in Blueberry and Their Regulatory Mechanism Controlling Chlorophyll Accumulation

SQUAMOSA Promoter Binding Protein (SBP) family genes act as central players to regulate plant growth and development with functional redundancy and specificity. Addressing the diversity of the SBP family in crops is of great significance to precisely utilize them to improve agronomic traits. Blueberry is an important economic berry crop. However, the SBP family has not been described in blueberry. In the present study, twenty VcSBP genes were identified through data mining against blueberry transcriptome databases. These VcSBPs could be clustered into eight groups, and the gene structures and motif compositions are divergent among the groups and similar within each group. The VcSBPs were differentially expressed in various tissues. Intriguingly, 10 VcSBPs were highly expressed at green fruit stages and dramatically decreased at the onset of fruit ripening, implying that they are important regulators during early fruit development. Computational analysis showed that 10 VcSBPs were targeted by miR156, and four of them were further verified by degradome sequencing. Moreover, their functional diversity was studied in Arabidopsis. Noticeably, three VcSBPs significantly increased chlorophyll accumulation, and qRT-PCR analysis indicated that VcSBP13a in Arabidopsis enhanced the expression of chlorophyll biosynthetic genes such as AtDVR, AtPORA, AtPORB, AtPORC, and AtCAO. Finally, the targets of VcSBPs were computationally identified in blueberry, and the Y1H assay showed that VcSBP13a could physically bind to the promoter region of the chlorophyll-associated gene VcLHCB1. Our findings provided an overall framework for individually understanding the characteristics and functions of the SBP family in blueberry.

Genome-wide Association Analysis Reveals a Novel QTL CsFS1 for Fruit Skin Color in Cucumber

Fruit skin color is a crucial external trait that affects consumer preference in cucumber. In this study, an F2 population was constructed from a cross between the inbred lines G35 (with light-green fruit skin) and Q51 (with dark-green fruit skin) and used to investigate the inheritance patterns of fruit skin color in cucumber. Genetic analysis showed that dark-green fruit skin was dominant to light-green skin. A major QTL, Fruit skin 1 (CsFS1), was identified between 36.62 Mb and 39.77 Mb on chromosome 3 by BSA-seq and GWAS. We further narrowed down the CsFS1 locus to a 94-kb interval containing 15 candidate genes in three F2 recombinant individuals with light-green skin color and one BC4F2 recombinant individual with dark-green skin color. Among these genes, Csa3G912920, which encodes a GATA transcription factor, was expressed at a higher level in the pericarp of the NIL-1334 line (with light-green fruit skin) than in that of the NIL-1325 line (with dark-green fruit skin). This study provides a novel allele for the improvement of fruit skin color in cucumber breeding.

Does fruiting habit traits affect green fruit yield and its component traits in chilli (Capsicum annuum L.)?

In chilli, farmer preferences for fruiting habit traits, namely fruit orientation (pendant erect) and fruits node−1 (solitary cluster), vary from region to region. For increased acceptability by farmers/consumers, cultivars should be bred with preferred fruiting habit traits in high green fruit yielding background. An investigation was carried out to study the influence of fruiting habit traits on green fruit yield and its component traits during the 2016 and 2017 rainy seasons. The F2, backcross populations and near-isogenic lines (NILs) derived from crosses involving parents differing for single or both the fruiting habit traits were grouped into fruiting habit classes (single and cluster fruiting; and clustered erect, clustered pendant, single erect and single pendant). Significance of differences in fruit yield and its component traits between/among different fruiting habit trait classes were examined using ‘t’/‘F’ tests. The significance of ‘t’/‘F’ tests was regarded as pieces of evidence for considerable effects of fruiting habit traits on fruit yield and its component traits. While results-based on F2 and backcross generations indicated a lack of effects, those based on NILs indicated significant effects of fruiting habit traits on fruit yield and its component traits. The plants bearing pendant fruits (irrespective of whether they are clustered or single) produced a large number of heavier and longer fruits than those bearing erect fruits. The results are discussed about the strategies to develop high yielding cultivars with farmer preferred fruiting habit traits.

Induced defense response in red mango fruit against Colletotrichum gloeosporioides

Mango fruit exposed to sunlight develops red skin and are more resistant to biotic and abiotic stresses. Here we show that harvested red mango fruit that was exposed to sunlight at the orchard is more resistant than green fruit to Colletotrichum gloeosporioides. LCMS analysis showed high amounts of antifungal compounds, as glycosylated flavonols, glycosylated anthocyanins, and mangiferin in red vs. green mango skin, correlated with higher antioxidant and lower ROS. However, also the green side of red mango fruit that has low levels of flavonoids was resistant, indicated induced resistance. Transcriptomes of red and green fruit inoculated on their red and green sides with C. gloeosporioides were analyzed. Overall, in red fruit skin, 2,187 genes were upregulated in response to C. gloeosporioides. On the green side of red mango, upregulation of 22 transcription factors and 33 signaling-related transcripts indicated induced resistance. The RNA-Seq analysis suggests that resistance of the whole red fruit involved upregulation of ethylene, brassinosteroid, and phenylpropanoid pathways. To conclude, red fruit resistance to fungal pathogen was related to both flavonoid toxicity and primed resistance of fruit that was exposed to light at the orchard.

False Seedbeds Reduce Labor Requirements for Weeding in Chile Pepper

This study evaluated false seedbeds, which are sequences of irrigation and tillage that eliminate weed seedlings before crop planting, to reduce requirements for hand hoeing in chile pepper (Capsicum annuum). To address this objective, a field study was conducted near Las Cruces, NM from July 2015 to Oct. 2016 (experimental run 1) and July 2016 to Oct. 2017 (experimental run 2). False seedbeds were designed to target weeds that typically emerge after chile pepper planting. This was done by implementing false seedbeds the summer before chile pepper seeding. During chile pepper seasons, data included repeated measures of weed seedling emergence, amounts of time required for individuals to hoe field sections (i.e., hoeing time), and yields of two chile products: early harvest of green fruit and late harvest of red fruit. Hoeing time and yield data were included in cost–benefit analyses that also incorporated expenses and revenues projected by crop budget models for the study region. Results indicated false seedbeds caused a 54% decrease in weed population density during the chile pepper season of experimental run 1; however, for experimental run 2, false seedbeds did not affect cumulative weed seedling emergence. For both experimental runs, false seedbeds reduced hoeing times, suggesting that false seedbeds affected hoeing by means other than reduced weed density. After accounting for costs for implementation, false seedbeds reduced hand hoeing costs by $262/acre to $440/acre. These reductions in hoeing costs coincided with improved profitability in all but one combination of year and product. Green fruit yield was lower in false seedbed plots in experimental run 1; however, false seedbeds did not affect green fruit yield in experimental run 2, or red fruit yield in both experimental runs. These results indicate that false seedbeds implemented the summer before planting are promising techniques for reducing labor requirements for weeding in chile pepper production.