VaSDC1 Is Involved in Modulation of Flavonoid Metabolic Pathways in Black and Red Seed Coats in Adzuki Bean (Vigna angularis L.)

Seed coat colour is an important nutritional quality trait. Variations in anthocyanins and flavonoids induce the diversity of seed coat colour in adzuki bean (Vigna angularis L.). Red seed coat and black seed coat are important adzuki bean cultivars. Insights into the differences of flavonoid metabolic pathways between black and red adzuki bean are significant. In this study, we explored that the difference in seed coat colour between the red (Jingnong6) and the black (AG118) is caused by the accumulation of anthocyanins. The RNA-sequencing (RNA-Seq) and real-time reverse transcription (qRT)-PCR results showed that the Vigna angularis L. seed coat color (VaSDC1) gene, an R2R3-MYB transcription factor, should be the key gene to regulate the black and red seed coat colours. In three different colouring staes of seed development, VaSDC1 was specifically expressed in the black seed coat (AG118) landrace, which activates the structural genes of flavonoid metabolic pathways. As a result, this caused a substantial accumulation of anthocyanins and created a dark blue-black colour. In the red (Jingnong6) seed coat variety, low expression levels of VaSDC1 resulted in a lower accumulation of anthocyanins than in AG118. In addition, VaSDC1 was genetically mapped in the interval between simple-sequence repeat (SSR) markers Sca326-12, Sca326-4, and BAgs007 on chromosome 3 using an F4 segregating population derived from the cross between Jingnong6 and AG118. These results will facilitate the improvement of nutritional quality breeding in adzuki beans.

HISTOCHEMICAL INVESTIGATION ON ARCHIDENDRON BUBALINUM (JACK) NIELSEN.) FROM LAMPUNG, SUMATERA, INDONESIA

Objective: This study was to describe the histochemical and morpho-anatomical of kabau seeds originating from Lampung, Sumatra Indonesia. Methods: Microscopic anatomical analysis of kabau seeds was carried out on the parts of kabau seeds with an incision as thick as 100 μm. The sample was placed on a glass object and aquadest, glycerin and choral hydrate were added and then covered with a glass cover, then observed under the light microscope equipped with digital camera, and analysis using the S-Viewer program. Histochemical tests are carried out with cross sections, which are colored with the following: Lugol iodine solution; ferric chloride; dragendr of; ninhydrin; K2Cr2O7. Results: Macroscopic characteristics, neatly arranged cylindrical kabau seeds consisting of five to six seeds on each pod. Yellowish-white kabau seeds are covered in a black seed coat, have a distinctive odor like jengkol or jering, have a slightly bittersweetness and a soft texture. The size of kabau seeds is 2 cm in length and 1.5 cm in diameter. Microscopic results on kabau seeds, an incision in choral hydrate showed visible parts of the epicarpium, pericarpium contained oil sacs and cell nuclei, and endosperm in each part of the sac contained starch grains and oil sac bags that gave off odors to the head, incisions in the drops of aqua dest almost the same as choral hydrate except that the starch grains are more clearly visible and an average diameter of 5,176 μm starch can be calculated. Conclusion: Histochemical reaction in the Kabau seed incision gave positive results on tannins in the endosperm, positive results for amino acids in the endosperm of purple rice, positive for alkaloids in the epicardium and pericarpium parts; black color throughout the epicardium, pericarpium and endosperm indicates a lot of starch is contained; and there are polyphenols in the endosperm oil sac.

Breeding of Black Soybean with Green Cotyledon and Four Recessive Alleles for Lipoxygenase, Kunitz Trypsin Inhibitor, Lectin, and Stachyose

Anthocyanins from the black soybean seed coat are known to have many pharmaceutical effects. However, black soybean seed contains antinutritional factors such as lipoxygenase, Kunitz trypsin inhibitor (KTI), lectin, and stachyose. The genetic removal of these components will improve the nutritional value of black soybean seed. The objective of this research was to breed a soybean strain with the black seed coat color, green cotyledon color, and tetra recessive allele (lox1lox2lox3/lox1lox2lox3-ti/ti-le/le-rs2/rs2) for lipoxygenase, KTI, lectin, and stachyose components. Eight parents were used to breed the tetra null strain. Analysis of lipoxygenase, KTI, lectin, and stachyose components in mature seeds was conducted by SDS-PAGE, Western blot, and HPLC. The soybean line with the black seed coat color, the green cotyledon color, a large seed size, and tetra recessive alleles has purple flowers, a determinate growth habit, and brown pods at maturity. The stem height of the breeding line was 52.3 cm. The 100-seed weight of the breeding line was 35.2 g and the yield (Ton/ha) was 2.50. The stachyose content of the breeding line was 3.30 g/kg. This is the first soybean strain with the black seed coat color, the green cotyledon color, a large seed size, and tetra null alleles (lox1lox2lox3/lox1lox2lox3-ti/ti-le/le-rs2/rs2, low content of stachyose, free of lipoxygenase, KTI, and lectin proteins).

The inhibitory effects of polyphenolic compounds on the damage caused by safflower fly (Acanthiophilus helianthi) in Carthamus spp.

Seed coat color is probably a determinant factor in the antibiosis mechanism of developing resistance to safflower fly (Acanthiophilus helianthi). The purpose of this study was to determine the relationship between the phytochemical content of safflower (Carthamus spp.) seed coat extract with the seed coat color and the damage caused by safflower fly. To this end, germplasm consisting of the cultivated species (C111, with a white seed coat), the wild species (Glaucus and lanatus with a black seed coat; Azar with a brown seed coat), and a breeding line (A82 with a black seed coat) was formed. After cultivating the genotypes, the seed loss (%) and its relationship with the polyphenolic compounds and Cyanidin-3-glucoside of the seed coat extract were examined. Agricultural and phytochemical trait data were analyzed through a completely random block design. With a significant difference from other samples, a minimum damage percentage was observed in Lanatus, Glaucus, and A82 genotypes with a black coat (P < 0.05). The concentration of phenolic compounds, that is, chlorogenic acid, caffeic acid, and p-coumaric acid, except for ferulic acid, was almost equal in all genotypes. However, there was an inverse and direct relationship between the concentration of four polyphenolic compounds (rutin, apigenin, quercetin, and ferulic acid) and Cyd-3-glu content with resistance safflower fly, respectively. In general, flavonoid compounds, that is, rutin, quercetin, and apigenin, affect the resistance probably through antibiosis mechanism so that there was a negative relationship between the concentration of these compounds and resistance to safflower fly.

Inheritance of black seed coat colour in lentil

The purpose of the research was to determine the inheritance of black seed coat colour in lentil variety ‘Beluga’. The seeds collected from F1 plants in cross of ‘Rauza’ (yellow seeds) × ‘Beluga’ (black seeds) were of grey colour with black mottles. F2 ratio of nonblack and black seeds was 62:6, which corresponded to 15:1 dihybrid segregation. It is concluded that the black seed coat colour of ‘Beluga’ is controlled by two dominant genes.

Identification of candidate genes controlling black seed coat and pod tip color in cowpea (Vigna unguiculata L. Walp)

ABSTRACTSeed coat color is an important part of consumer preferences for cowpea (Vigna unguiculata L. Walp). Color has been studied in numerous crop species and has often been linked to loci controlling the anthocyanin biosynthesis pathway. This study makes use of available resources, including mapping populations, a reference genome, and a high-density single nucleotide polymorphism genotyping platform, to map the black seed coat and purple pod tip color traits in cowpea. Several gene models encoding MYB domain protein 113 were identified as candidate genes. MYB domain proteins have been shown in other species to control expression of genes encoding enzymes for the final steps in the anthocyanin biosynthesis pathway. PCR analysis indicated that a presence/absence variation of one or more MYB113 genes may control the presence or absence of black pigment. A PCR marker has been developed for black seed coat color in cowpea.