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G-protein from Medicago sativa: functional association to photoreceptors.

ABSTRACT: G-protein subunits were characterized from Medicago sativa (alfalfa) seedlings. Crude membranes and GTP-Sepharose-purified fractions were electrophoresed on SDS/polyacrylamide gels and analysed by Western blotting with 9193 (anti-alpha common) and AS/7 (anti-alpha t, anti-alpha i1 and anti-alpha i2) polyclonal antibodies. These procedures led to the identification of a specific polypeptide band of about 43 kDa. Another polypeptide reacting with the SW/1 (anti-beta) antibody, of about 37 kDa, was also detected. The 43 kDa polypeptide bound specifically [alpha-32P]GTP by a photoaffinity reaction and was ADP-ribosylated by activated cholera toxin, but not by pertussis toxin. Irradiation of etiolated Medicago sativa protoplast preparations at 660 nm for 1 min produced a maximal increase in the guanosine 5'-[gamma-thio]triphosphate (GTP[35S])-binding rate. After this period of irradiation, the binding rate tended to decrease. The effect of a red-light (660 nm) pulse on the binding rate was reversed when it was immediately followed by a period of far-red (> 730 nm) illumination. These results may suggest that activation of GTP[S]-binding rate was a consequence of conversion of phytochrome Pr into the Ptr form.

SUBMITTER: Muschietti JP

PROVIDER: S-EPMC1132536 | biostudies-other | 1993 Apr

REPOSITORIES: biostudies-other

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Project description:Genetic progress for forage quality has been poor in alfalfa (Medicago sativa L.), the most-grown forage legume worldwide. This study aimed at exploring opportunities for marker-assisted selection (MAS) and genomic selection of forage quality traits based on breeding values of parent plants. Some 154 genotypes from a broadly-based reference population were genotyped by genotyping-by-sequencing (GBS), and phenotyped for leaf-to-stem ratio, leaf and stem contents of protein, neutral detergent fiber (NDF) and acid detergent lignin (ADL), and leaf and stem NDF digestibility after 24 hours (NDFD), of their dense-planted half-sib progenies in three growing conditions (summer harvest, full irrigation; summer harvest, suspended irrigation; autumn harvest). Trait-marker analyses were performed on progeny values averaged over conditions, owing to modest germplasm × condition interaction. Genomic selection exploited 11,450 polymorphic SNP markers, whereas a subset of 8,494 M. truncatula-aligned markers were used for a genome-wide association study (GWAS). GWAS confirmed the polygenic control of quality traits and, in agreement with phenotypic correlations, indicated substantially different genetic control of a given trait in stems and leaves. It detected several SNPs in different annotated genes that were highly linked to stem protein content. Also, it identified a small genomic region on chromosome 8 with high concentration of annotated genes associated with leaf ADL, including one gene probably involved in the lignin pathway. Three genomic selection models, i.e., Ridge-regression BLUP, Bayes B and Bayesian Lasso, displayed similar prediction accuracy, whereas SVR-lin was less accurate. Accuracy values were moderate (0.3-0.4) for stem NDFD and leaf protein content, modest for leaf ADL and NDFD, and low to very low for the other traits. Along with previous results for the same germplasm set, this study indicates that GBS data can be exploited to improve both quality traits (by genomic selection or MAS) and forage yield.

Project description:This study explored the effects of four sugar source treatments, including no sugar (CON), fructose (FRU), pectin (PEC), and starch (STA), on the microbiota and metabolome of alfalfa (Medicago sativa) silage. The bacterial community was determined via 16S rRNA gene sequencing. The metabolome was analyzed using ultra high-performance liquid chromatography tandem time-of-flight mass spectrometry (UHPLC/TOF-MS). After 60 days of fermentation, the pH values in FRU and PEC were lower than those in STA and CON. FRU had a greater lactic acid concentration compared to STA and CON. Weissella (47.44%) and Lactobacillus (42.13%) were the dominant species in all four groups. The abundance of Pediococcus was lower, and the abundance of Leuconostoc, Pantoea, and Microbacterium was higher, in FRU compared to CON. The abundance of norank_f__Bacteroidales_S24-7_group was higher, and the abundance of Turicibacter was lower, in both FRU and PEC than in CON. Leuconostoc was negatively correlated with the pH value, and Pediococcus was positively correlated with the pH value. No microbiomes were detected as discriminative features between STA and CON. The addition of FRU and PEC presented more peptides, such as Leu-Val-Thr, Leu-Phe, Ile-Pro-Ile, Val-Trp, and Ile-Leu-Leu but a lower abundance of metabolites for triterpene glycosides including sanchinoside B1, medicagenic acid, betavulgaroside IV, and prosapogenin compared to CON. The addition of PEC presented more phenyllactic acid compared to CON. Our study demonstrated that the addition of pectin and fructose improved the quality of alfalfa silage mainly by promoting Leuconostoc, Pantoea, and Microbacterium, and inhibiting Pediococcus in FRU, and promoting norank_f__Bacteroidales_S24-7_group and inhibiting Turicibacter in both FRU and PEC; this was due to altered metabolic profiles resulting from antifungal activity and decreased triterpene glycoside accumulation. This study improves our understanding of ensiling mechanisms related to the contributions of sugar.

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G-protein from Medicago sativa: functional association to photoreceptors.

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