Population structure analysis and determination of neurotoxin content in a set of grass pea (Lathyrus sativus L.) accessions of Bangladesh origin

Grass pea(Lathyrus sativus L.) is a crop that is considered one of the more resilient to climate change. With protein-rich seeds and leaves, it has strong potential as human food as well as animal feed and fodder. However, genetic improvement in this crop remains stagnant owing to the poor characterization of its genetic resources. In this study, we characterized 118 accessions of grass pea with 18 EST-SSR markers. A total of 118 accessions, 101 of L. sativus(100 cultivated accessions from Bangladesh and one wild accession) and 17 wild accessions of other Lathyrus species, were used. A total of 67 alleles were detected, with an average of 3.72 alleles per locus and average polymorphism information content of 0.52. A dissimilarity matrix was formed and hierarchical cluster analysis performed using the UPGMA method grouped genotypes into four main clusters. Cluster analysis based on the genetic dissimilarity revealed a clear grouping of the 100 cultivated and 18 wild accessions into four main groups. Group I consisted of 20 accessions with high β-N-oxalyl-L-α,β-diaminopropionic acid(β-ODAP) concentration. Of these 20 accessions, 17 were wild accessions. Only one wild accession(L. cicera) was clustered in group II, which contained 35 accessions. Most of the group II accessions contained low β-ODAP. Group III was represented by 34 accessions, many of them with high β-ODAP. Group IV consisted of 29 accessions, a few of which had very high β-ODAP concentrations. Analysis of molecular variance of the microsatellite data showed significantly higher values of molecular variance between(83%) than within(17%) populations. These characterized accessions will be useful in grass pea breeding programs.

Determination of Neurotoxin β-ODAP and Non-protein Amino Acids in Lathyrus Sativus by High-Performance Liquid Chromatography with Precolumn Derivatization with 6-Amino quinolyl-N-hydroxysuccinimidyl Carbamate (AQC)

A new method was developed for the quantitative determination of the neurotoxic nonprotein amino acid, 3-N-oxalyl-L-2,3-diaminopropioni acid (β-ODAP), its nontoxic α-isomer and other non-protein amino acids in the plant samples of Lathyrus sativus after derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) by reversed-phase high-performance liquid chromatography (HPLC). 2-Amino butyric acid (ABA) was used as an internal standard.The RP HPLC detection limit for both isomers is 1.8 ng with good response linearity. The results are compared with a colorimetric method.

HPLC Determination of Neurotoxin β-N-oxalyl-L-α, β-diaminopropionic acid and Its α -Isomer in Lathyrus sativus by Precolumn Derivatisation with 1-Fluoro-2,4-dinitrobenzcne

A rapid and simple method is presented for determining β-N-oxalyl-α. β- diaminopropionic acid (β -ODAP) and its much less toxic α -isomer (α -ODAP) in Lathyrus sativus. Seed and foliage extracts of Lathyrus sativus were treated with 1-fluoro-2,4-dinitrobenzene (FDNB) and a reversed-phase high-performance liquid chromatographic method for the separation of the derivatives in the pmol range is reported.

Effect of Dietary Administration of Lathyrus sativus Pulse on Intestinal Biochemical Parameters in Normal and Scorbutic Guinea Pigs

Objective In order to investigate that ascorbic acid deficiency is responcible for lathyrus toxicity, the effect of dietary feeding of lathyrus pulse in normal and scorbutic guinea pigs for 3 months, on intestinal biochemical parameters was undertaken. Methods The intestinal brush border membrane (BBM) marker and xenobiotic metabolising enzymes (XME) were assayed. Results Exposure to 80% lathyrus alone and in scorbutic conditions showed significant inhibition of alkaline phosphatase (28%-30%), sucrase (19%) and γ-glutamyl transpeptidase (GGT) (15%-27%) enzymes, while Ca2+-Mg2+-ATPase was significantly inhibited (38%) in scorbutic plus lathyrus treated group. The phase I XME (AHH) remained unchanged while the phase II enzyme glutathione-S-tranferase (GST) was significantly decreased (20%-22%) in lathyrus and scorbutic plus lathyrus treated groups. Quinone reductase (QR) activity was found to be significantly decreased in lathyrus exposed group (20%). The intestinal biomarker contents including hexose (25%-34%) and phospholipids (20%-40%) were significantly reduced in lathyrus and scorbutic plus lathyrus exposed animals, while sialic acid showed a significant decrease (28%) in scorbutic plus lathyrus treated group. However, cholesterol levels were signifcantly enhanced (15%-28%) in lathyrus and scorbutic plus lathyrus treated animals. Conclusion The results indicate that oral feeding of lathyrus pulse to guinea pigs can alter BBM parameters as well as XME, which may result in the intestinal toxicity. Further, ascorbic acid deficiency could be one of the pre-disposing factors of lathyrus toxicity.

L-Homoarginine Accumulation in Grass Pea (<i>Lathyrus sativus</i>L.) Dry Seeds. A Preliminary Survey

Grass pea (Lathyrus sativus L.) has great agronomic potential as grain and forage legume, and presently is con-sidered as a model crop for sustainable agriculture. However, the development into an important food legume has been hin- dered by the presence of the neurotoxic amino acid β-N-Oxalyl-α, β-diaminopropionic acid (β-ODAP). Recent studies reported that homoarginine (Har) can counteract this toxic action. This research was undertaken to shed light on the variation of Har amount within grass pea. The influence of the environment and of the year-to-year variation of cli- matic conditions was also investigated. Seven Italian grass pea ecotypes were evaluated for two subsequent growing seasons in two locations of southern Italy. In contrast with previous studies collected data evidenced a significant variation of Har amount among the tested ecotypes. Moreover, a significant positive correlation between Har and ODAP level was observed. The effect of year-to-year variation of temperature and rainfall quantity is also discussed.

Novel associations between rhizobial populations and legume species within the genera Lathyrus and Oxytropis grown in the temperate region of China

Fifty rhizobial isolates of Lathyrus and Oxytropis collected from northern regions of China were studied in their genotypic characterization based upon analyses of ARDRA, 16S-23S IGS PCR-RFLP, TP-RAPD, MLEE, sequences of 16S rDNA gene and housekeeping genes of atpD, recA and glnII. The results demonstrated that most of the Lathyrus rhizobia belonged to Rhizobium and most of the Oxytropis rhizobia belonged to Sinorhizobium. A novel group of Rhizobium sp. I and S. meliloti were identified as the main microsymbionts respectively associated with Lathyrus and Oxytropis species in the collection area, which were new associations between rhizobia and the mentioned hosts. This study also provides new evidence for biogeography of rhizobia.

Performance index: An expeditious tool to screen for improved drought resistance in the Lathyrus genus

Some species of the Lathyrus genus are among the most promising crops for marginal lands, with high resilience to drought, flood, and fungal diseases, combined with high yields and seed nutritional value. However, lack of knowledge on the mechanisms underlying its outstanding performance and methodologies to identify elite genotypes has hampered its proper use in breeding. Chlorophyl a fast fluorescence transient （JIP test）, was used to evaluate water deficit （WD） resistance in Lathyrus genus. Our results reveal unaltered photochemical values for al studied genotypes showing resistance to mild WD. Under severe WD, two Lathyrus sativus genotypes showed remarkable resilience maintaining the photochemical efficiency, contrary to other genotypes studied. Performance index （PIABS） is the best parameter to screen genotypes with improved performance and grain production under WD. Moreover, we found that JIP indices are good indicators of genotypic grain production under WD. Quantum yield of electron transport （wEo） and efficiency with which＆trapped excitons can move electrons further than QA （c0） revealed as important traits related to improved photosyn-thetic performance and should be exploited in future Lathyrus germplasm improvements. The JIP test herein described showed to be an expeditious tool to screen and to identify elite genotypes with improved drought resistance.

Effect of water regimes and organic matters on transport of arsenic in summer rice (Oryza sativa L.)

The arsenic contamination in soil-water-plant systems is a major concern of where, the groundwater is being contaminated with arsenic （above 0.01 mg/L） in the Indian subcontinent. The study was conducted with organic matter to find out the reducing effect on arsenic load to rice （cv. Khitish）. It was observed that intermittent ponding reduced arsenic uptake （23.33% in root, 13.84% in shoot and 19.84% in leaf） at panicle initiation stage, instead of continuous ponding. A decreasing trend of arsenic accumulation （root straw husk whole grain milled grain） was observed in different plant parts at harvest. Combined applications of lathyrus ＋ vermicompost ＋ poultry manure reduced arsenic transport in plant parts （root, straw, husk, whole grains and milled grain） which was significantly at par （p 0.05） with chopped rice straw （5 tons/ha ） ＋ lathyrus green manuring （5 tons/ha） in comparison to control and corresponding soils. A significant negative correlation of arsenic with phosphorus （grain P with arsenic in different parts R 2 = 0.627–0.726 at p 0.01） was observed. Similarly, soil arsenic had a negative correlation with soil available phosphorus （R 2 = 0.822 at p 0.001） followed by soil nitrogen （R 2 = 0.762 at p 0.01） and soil potassium （R 2 = 0.626 at p 0.01）. Hence, effective management of contaminated irrigation water along with organic matter could reduce the arsenic build up to plants and soil.

不同钠盐胁迫对松柳种子萌发及幼苗生长的影响

为了研究不同钠盐胁迫对松柳种子萌发及幼苗生长影响,采用0~400 mmol/L的NaCl、0~100 mmol/L的Na2CO3和0~100 mmol/L的NaHCO3溶液处理松柳(Lathyrus quinquenervius)种子,每种溶液设置6个浓度梯度,以此研究盐胁迫对种子萌发和幼苗生长的影响。研究结果表明:NaCl浓度在0~100 mmol/L时对种子发芽的各项指标影响不明显,在浓度大于200 mmol/L时,显著降低了种子发芽率、相对发芽率、发芽势和发芽指数,盐害率显著升高;Na2CO3溶液在所有浓度下均显著降低种子发芽指标,盐害率显著升高,仅在2 mmol/L时未影响种子发芽率;NaHCO3溶液在浓度为0~20 mmol/L时对种子发芽的各项指标无显著影响,而浓度>20 mmol/L后,对种子的发芽势、发芽指数、耐盐指数均产生显著抑制,浓度100 mmol/L时,盐害率显著升高。三种盐胁迫对苗高生长均表现为低浓度显著促进,高浓度显著抑制,NaCl和Na2CO3对胚根生长表现为低浓度无显著影响而高浓度显著抑制,NaHCO3对胚根生长以及三种盐对胚芽生长则始终呈抑制作用。