Methylation and bio-accessibility assessment of arsenate in crickets (Gryllusbimaculatus).

The ability of an organism to biomethylate toxic inorganic arsenic (As) determines both, the amount of As available for uptake higher up the food chain and the toxicity of bioavailable As. An exposure study was conducted to determine ability of farmed crickets to metabolize dietary arsenate. Crickets were exposed to 1.3 ± 0.1, 5.1 ± 2.5 and 36.3 ± 5.6 mg kg-1 dietary arsenate and quantitation of total As showed retention of 0.416 ± 0.003, 1.3 ± 0.04 and 2.46 ± 0.09 mg kg-1, respectively. Speciation analysis revealed that crickets have well developed ability to biomethylate dietary arsenate and the most abundant methylated As compound was DMA followed by MMA, TMAO and an unknown compound. Arsenobetaine, although present in all feed, control and As-rich, was measured only in the control crickets. To assess the bio-accessibility of the As species, crickets were subjected to simulated gastrointestinal digestion. The results showed that majority of As was extracted in saliva, followed by gastric and intestinal juice, which mass fraction was equal to residue. Over 78% of total As was shown to be bio-accessible with methylated species reaching 100% and iAs over 79% bio-accessibility. Additionally, arsenite and arsenate have shown different distributions between sequential leachate solutions. Bioaccumulation of As was observed in the studied crickets although it does not seem to occur to the same extent at higher exposure levels.

Mechanisms of Trichomes and Terpene Compounds in Indigenous and Commercial Thai Rice Varieties against Brown Planthopper.

Plant trichomes generally act as a physical defense against herbivore attacks and are present in a variety of plants, including rice plants. This research examined the physical and chemical defenses of rice plants against the brown planthopper (BPH), Nilaparvata lugens (Stål) (Hemiptera: Delphacidae). A total of 10 rice varieties were used in this study. An electron microscope was used to observe trichomes. Constitutive and induced volatile compound profiles were assessed using GC-MS analyses. The preference of BPH for volatiles from the 10 rice plants was tested using a two-choice arena olfactometer system. The density of prickle trichomes had a negative relationship with the BPH injury level. Without BPH infestation, the volatile of the most resistant rice variety (Rathu Heenati (RH)) was preferred by BPH than those of the other varieties, with the exception of Gled Plah Chawn. However, the relative BPH preference for volatiles from the RH variety decreased during BPH infestation. When rice plants were infested by BPH, the numbers of VOCs and these quantities decreased. In the RH variety, the emission of essentities found without BPH infestation ceased during infestation by BPH. During the BPH infestation, rice plants started to emit new VOCs that were not detected before the BPH infestation started. In conclusion, we discovered that rice plants defended against BPH by changing VOC components during BPH infestation and ß-Sesquiphellandrene was likely the most effective component.

Rice Sesquiterpene Plays Important Roles in Antixenosis against Brown Planthopper in Rice.

The rice sesquiterpene synthase II gene (OsSTPS2, LOC_Os04g27430), which is involved in the antixenosis defense mechanism of rice against brown planthopper (BPH) infestation, was identified in the BPH-resistant rice variety Rathu Heenati (RH). In contrast, the gene was not functional in the BPH-susceptible rice variety KDML105 (KD). Single-nucleotide polymorphisms (SNPs) in the promoter region and in exon 5 of the gene and a seven amino acid deletion in the deduced protein sequence are suggested as factors that negatively regulate the function of the gene. Sequence analysis of the promoter region and expression analysis of the OsSTPS2 gene in several rice genotypes revealed the correlation of SNPs of the ATHB-1, SBE1, and P-factor with the expression of the gene. Genomic and complementary DNA (cDNA) sequence analysis at exon 5 of the gene showed that the 21 bp deletion naturally occurred in several rice genotypes. The antixenosis of the BPH feeding preference (AFP) of rice varieties differed in the seven amino acid deletion lesion of the gene, suggesting that the seven amino acid deletion negatively controls the antixenosis mechanism during BPH infestation. Analysis of the plant volatile compounds released after BPH infestation suggested that E-ß-farnesene (EBF) is the major product of the OsSTPS2 gene.

Pseudo-backcrossing design for rapidly pyramiding multiple traits into a preferential rice variety.

BACKGROUND: Pyramiding multiple genes into a desirable genetic background can take years to accomplish. In this paper, a pseudo-backcrossing scheme was designed to shorten the backcrossing cycle needed. PinK3, an aromatic and potentially high-yielding rice variety-although one that is intolerant to flash flooding (Sub) and susceptible to bacterial leaf blight (BB), leaf-neck blast (BL) and the brown planthopper (BPH)-was used as a genetic basis for significant improvements through gene pyramiding. RESULTS: Four resistance donors with five target genes (Sub1A-C, xa5, Xa21, TPS and SSIIa) and three QTLs (qBph3, qBL1 and qBL11) were backcrossed individually using markers into the pseudo-recurrent parent 'PinK3' via one cycle of backcrossing followed by two cycles of pseudo-backcrossing and three selfings with rigorous foreground marker-assisted selection. In total, 29 pseudo-backcross inbred lines (BILs) were developed. Genome composition was surveyed using 61 simple sequence repeats (SSRs), 35 of which were located on six carrier chromosomes, with the remainder located on six non-carrier chromosomes. The recurrent genome content (%RGC) and donor genome content (%DGC), which were based on the physical positions of BC1F2, ranged from 69.99 to 88.98% and 11.02 to 30.01%, respectively. For the pseudo-BC3F3BILs, the %RGC and %DGC ranged from 74.50 to 81.30% and 18.70 to 25.50%, respectively. These results indicated that without direct background selection, no further increases in %RGC were obtained during pseudo-backcrossing, whereas rigorous foreground marker-assisted selection tended to reduce linkage drag during pseudo-backcrossing. The evaluation of new traits in selected pseudo-BC3F3BILs indicated significant improvements in resistance to BB, BL, BPH and Sub compared with PinK3, as well as significant improvements in grain yield (21-68%) over the donors, although yield was 7-26% lower than in 'PinK3'. All pyramided lines were aromatic and exhibited improved starch profiles, rendering them suitable for industrial food applications. CONCLUSIONS: Results show that our new pyramiding platform, which is based on marker-assisted pseudo-backcrossing, can fix five target genes and three QTLs into a high-yielding pseudo-recurrent background within seven breeding cycles in four years. This multiple pseudo-backcrossing platform decreases the time required to generate new rice varieties exhibiting complex, durable resistance to biotic and abiotic stresses in backgrounds with desirable qualities.

Single-feature polymorphism mapping of isogenic rice lines identifies the influence of terpene synthase on brown planthopper feeding preferences.

BACKGROUND: Bph3, a major brown planthopper (BPH) resistance locus derived from the rice cultivar Rathu Heenati (RH), has been used as a stable donor of traits that improve highly susceptible aromatic rice varieties in Thailand. Map-based cloning was initiated using a set of isogenic lines (ILs) harboring the major Bph3 locus on chromosome 6. IL genomes were scanned with a 57 K Affymetrix Rice GeneChip to identify the gene responsible for Bph3. FINDINGS: Single-feature polymorphism (SFP) mapping was used to localize 84 candidate genes. An expression analysis of 15 selected candidate genes in the aromatic rice cultivar KDML105 (KD) and the ILs under normal conditions revealed two differentially expressed sequences. Following hopper feeding, only one candidate gene, Os04g27430, was differentially expressed. Os04g27430 encodes a putative sesquiterpene synthase (STPS) gene that was induced by BPH feeding in ILs. An antixenosis test in three selected ILs revealed a major role for STPS in insect preference during the first 120 hours of the rice-insect interaction. Functional SNPs in exon 5 that resulted in the deletion of seven amino acids in the susceptible rice line were identified. Moreover, three additional SNPs associated with three transcription binding sites were also identified, which might explain the differential response of Os04g27430 during the anti-feeding test. CONCLUSION: Os04g27430 is the second known rice STPS induced by BPH. The gene may involve an antixenosis BPH resistance mechanism. The combination of the STPS and the Bph3 locus was more effective than Bph3 alone in the tested ILs.