Nitrogen source-dependent inhibition of yeast growth by glycine and its N-methylated derivatives

The effect of nitrogen source on the inhibitory properties of glycine and its N-methylated derivatives N-methylglycine (sarcosine), N,N-dimethylglycine, N,N,N-trimethylglycine (glycine betaine) on yeast growth was investigated. On solid minimal medium, all four glycine species completely or partially inhibited growth of Kluyveromyces lactis, Komagataella pastoris, Ogataea arabinofermentans, Spathaspora passalidarum and Yamadazyma tenuis at concentrations 5–10 mM when 10 mM NH4Cl was the sole source of nitrogen. If NH4Cl was substituted by sodium L-glutamate as the sole source of nitrogen, obvious growth inhibition by glycine and its N-methylated derivatives was generally not observed in any of these species. No obvious growth inhibition by any of the glycine species at a concentration of 10 mM was observed in Cyberlindnera jadinii, Lipomyces starkeyi, Lodderomyces elongisporus, Scheffersomyces stipitis or Yarrowia lipolytica on solid minimal medium irrespective of whether the nitrogen source was NH4Cl or sodium L-glutamate. Growth inhibition assays of K. pastoris in liquid minimal medium supplemented with increasing concentrations of N,N-dimethylglycine demonstrated inhibitory effects for nine tested nitrogen sources. In most cases, N,N-dimethylglycine supplementation caused a decrease in growth efficiency that appeared to be proportional to the concentration of N,N-dimethylglycine. The biological relevance of these results is discussed.

Flavonoids from three Wild Glycine Species in Japan and Taiwan

Fourteen flavonols, four flavones and six isoflavones were isolated from the aerial parts of two Japanese Glycine species, G. tabacina and G. koidzumii, and the leaves of Taiwanese G. max subsp. formosana. Of their flavonoids, twelve flavonols were identified as kaempferol 3- O-sophoroside (1), 3- O-rutinoside (2), 3- O-robinobioside (3) and 3- O-rhamnosyl-(1→4)-[rhamnosyl-(1→6)-galactoside] (4), quercetin 3- O-gentiobioside (5), 3- O-glucoside (6), 3- O-galactoside (7), 3- O-rutinoside (8), 3- O-robinobioside (9) and 3- O-rhamnosyl-(1→4)-[rhamnosyl-(1→6)-galactoside] (10), and isorhamnetin 3- O-rutinoside (11) and 3- O-robinobioside (12). Other two flavonols were characterized as isorhamnetin 3- O-rhamnosylrhamnosylglucoside (13) and 3- O-rhamnosylrhamnosylgalactoside (14). Four flavones and six isoflavones were estimated as schaftoside (15), apigenin 6,8-di- C-arabinoside (16), luteolin 7- O-glucoside (17) and chrysoeriol 7- O-glucoside (18), and daidzein 7- O-glucoside (19), 4'- O-glucoside (20) and 7- O-xylosylglucoside (21), genistein 7- O-glucoside (22) and 4'- O-glucoside (23), and 3'- O-methylorobol 7- O-glucoside (24). Although flavonoid composition of G. tabacina and G. koidzumii was similar to each other, that of G. max subsp. formosana was different with those of two Japanese Glycine species described above. Flavonoids of their Glycine species were reported for the first time except for those of G. tabacina.

Accessions of Perennial Glycine Species With Resistance to Multiple Types of Soybean Cyst Nematode (Heterodera glycines)

Soybean cyst nematode (SCN; Heterodera glycines; HG) is a widely occurring and damaging pathogen that limits soybean production. Developing resistant cultivars is the most cost-effective method for managing this disease. Genes conferring SCN resistance in soybean have been identified; however, there are SCN populations that overcome known resistance genes. In order to identify additional sources of resistance and potentially new resistance genes, 223 plant introductions (PIs) of G. tomentella and 59 PIs of 12 other perennial Glycine species were inoculated with HG Types 0, HG 2, and HG 1.2.3, and then 36 PIs out of this set were further evaluated with HG Type 1.2.3.4.5.6.7, a population that overcomes all the resistance genes in soybean. Of 223 G. tomentella PIs evaluated, 86 were classified as resistant to three HG types, 69 as resistant to two HG types, and 22 as resistant to one HG type. Of the other 12 perennial Glycine species, all PIs of G. argyrea and G. pescadrensis were resistant to all three HG types. Of the 36 PIs challenged with HG Type 1.2.3.4.5.6.7, 35 were resistant with 16 showing no cyst reproduction. Our study confirms that there are high levels of resistance to SCN among the perennial Glycine species. This represents an untapped resource for use in genetic studies and for improving resistance to SCN in soybean.

Development of a Competent and Trouble Free DNA Isolation Protocol for Downstream Genetic Analyses in Glycine Species

Extraction of deoxyribose nucleic acid (DNA) from plants is preliminary step in molecular biology. Fast and cost effective genomic DNA isolation from Glycine species for downstream application is a major bottleneck. Here we report a high throughput and trouble free method for genomic DNA extraction from leaf and seeds of Glycine species with high quality and quantity. Protocol reports the optimization by employing different concentrations of CTAB and PVP in extraction buffer. Efficiency of optimized protocol was compared with frequently used DNA extraction methods. Wide adoptability and utility of this protocol was confirmed by DNA extraction from leaves as well as seeds of G. max, G. soja, G. tomentella and G. latifolia. Extracted DNA was successfully subjected to PCR amplification of five microsatellite markers and four putative glycosyltransferase genes. DNA extraction protocol is reproducible, trouble free, rapid and can be adopted for plant molecular biology applications.