A Study on the Activity of Carboxylesterase and the Differential Expression of Its Gene in the Midguts of Bombyx mori Resistant to BmDNV-Z

This study was to discuss the relationship among the change in the activity of Bombyx mori carboxylesterase （BmCarE） in the midguts, the differential expression of BmCarE gene （bmcare） in the midguts, and the ability of Bombyx mori resistant to densonucleosis virus （BmDNV）, and to elucidate the molecular mechanism of resistance to BmDNV-Z. With two silkworm strains, HUABA, which is susceptible to BmDNV-Z, and BC8 （a near isogenic line of HUABA）, which is completely resistant to the same virus, as materials, the activity of BmCarE in the midgut was determined by Bio-Tek Synergy, and the differential expression of bmcare between the two strains was investigated by real-time fluorescence quantitative PCR, both at 12, 36, and 72 h post oral inoculation of the two strains with virus （hereafter referred as inoculation）. While the activity of BmCarE in the midguts of BC8 inoculation group at 12 h post inoculation was higher than that in the BC8 control group, the HUABA inoculated group, and the HUABA control group by 3.28, 2.26, and 3.02 times, respectively, with the difference being highly significant （P 〈 0.01）, there was no statistical difference among the other groups. The relative expression level of bmcare in the midguts of BC8 inoculation group at 12 h post inoculation was higher than that in the BC8 control group, the HUABA inoculation group, and the HUABA control group by 17.714, 21.76, and 15.09 times, respectively, with the difference being highly significant （P 〈 0.01）, and there was no statistical difference among other groups. The elevation of BmCarE activity and expression level of bmcare in the resistant strain at 12 h post inoculation may relate to the resistant gene （nsd/nsd） and the stimulation of BmDNV-Z. The molecular basis for the elevation of BmCarE activity in the resistant strain BC8 may be the change in the expression level of bmcare.     

Investigation of Genetic Variation of B-G Gene with PCR-SSCP and RFLP in Chinese Indigenous Chickens

To reveal genetic variation of MHC B-G gene at Chinese native chickens, two PCR primer pairs were designed to hybridize specifically with conserved sequences surrounding hypervariable regions within the B-G gene and used to amplify two DNA fragments in ten Chinese indigenous chicken breeds and one introduced breed. The fragments were cloned and sequenced to assure that the expected sequences of chicken B-G gene were isolated. Of which the 189 bp fragment encompassing the most variable region within exon 2 of B-G gene was employed for PCR-SSCP assay, this method provided evidence for the presence of at least 56 B-G genotypes in the Chinese chickens sampled. It revealed a high degree of diversity in B-G genes of Chinese local breeds; particularly, high variation of B-G gene was confirmed with the presence of 48 B-G genotypes within Tibetan chicken population. Not only can the B-G genotypes be used to preliminarily screen new B-G alleles, but also they would be utilized to investigate MHC haplotypes and matched unrelated donors for bone marrow transplantation in immune researches. Another fragment of 401 bp size spanning over partial intron 1 and exon 2 of B-G gene was employed for PCR-RFLP analysis with two restriction enzymes of Msp Ⅰ and Tas Ⅰ in the breeds sampled. In this part of the gene, three novel SNPs were detected at the two restriction sites. It was more generally found the transition of two nucleotides of A294G and T295C occurred at Tas Ⅰ restriction site, and consequently led to a non-synonymous substitution of asparagine into serine at position 54 within the deduced amino acid sequence of immunoglobulin variable-region-like domain encoded by the exon 2 of B-G gene. It was observed at rare frequency that an alone mutation of A294G occurring at the site, which also caused the substitution of amino acid, asparagine 54-to-serine;and we haven't found only single mutation occurred at position 295 of the restriction site. At the Msp Ⅰ site, the transversion of G318C led to a non-synonymous substitution, glutamine 62-to-histidine. The variations at expression level caused by the genetic variability of B-G gene may bring about the changes in immune specificity of B-G antigen finally. Furthermore, two alleles, A and B, were identified at Msp Ⅰ and Tas Ⅰ loci of B-G gene, respectively. The allele frequencies were estimated, which gave a nonsymmetrical distribution either in the eight Chinese local breeds or in the introduced breed. By comparison, allele A at Msp Ⅰ locus was tended to be dominative, while, the allele B at Tas Ⅰlocus was tended to be prevalent in the breeds analyzed. It is concluded that the genetic variability of B-G gene revealed by the PCR-SSCP and RFLP assays in Chinese native chickens provide molecular data for further investigating the varied immune functions of B-G antigen; and the PCR-RFLPs at Msp Ⅰ and Tas Ⅰ loci of B-G gene might be used as genetic markers in selecting for the traits of disease resistance in chicken breeding.     

气候和土壤质地对土壤有机碳影响的区域差异研究

The agricultural soil carbon pool plays an important role in mitigating greenhouse gas emission ana unaerstanamg the son orgamc carbon-climate-soil texture relationship is of great significance for estimating cropland soil carbon pool responses to climate change. Using data from 900 soil profiles, obtained from the Second National Soil Survey of China, we investigated the soil organic carbon （SOC） depth distribution in relation to climate and soil texture under various climate regimes of the cold northeast region （NER） and the warmer Huang-Huai-Hai region （HHHR） of China. The results demonstrated that the SOC content was higher in NER than in HHHR. For both regions, the SOC content at all soil depths had significant negative relationships with mean annual temperature （MAT）, but was related to mean annual precipitation （MAP） just at the surface 0-20 cm. The climate effect on SOC content was more pronounced in NER than in HHHR. Regional differences in the effect of soil texture on SOC content were not found. However, the dominant texture factors were different. The effect of sand content on SOC was more pronounced than that of clay content in NER. Conversely, the effect of clay on SOC was more pronounced than sand in HHHR. Climate and soil texture jointly explained the greatest SOC variability of 49.0% （0-20 cm） and 33.5% （20-30 cm） in NER and HHHR, respectively. Moreover, regional differences occurred in the importance of climate vs. soil texture in explaining SOC variability. In NER, the SOC content of the shallow layers （0-30 cm） was mainly determined by climate factor, specifically MAT, but the SOC content of the deeper soil layers （30-100 cm） was more affected by texture factor, specifically sand content. In HHHR, all the SOC variability in all soil layers was predominantly best explained by clay content. Therefore, when temperature was colder, the climate effect became stronger and this trend was restricted by soil depth. The regional differences and soil depth influence underscored the importance of explicitly considering them in modeling long-term soil responses to climate change and predicting potential soil carbon sequestration.     

耕作方式对双季稻田土壤有机碳储量的影响

Tillage practices can potentially afect soil organic carbon (SOC) accumulation in agricultural soils. A 4-year experiment was conducted to identify the influence of tillage practices on SOC sequestration in a double-cropped rice (Oryza sativa L.) field in Hunan Province of China. Three tillage treatments, no-till (NT), conventional plow tillage(PT), and rotary tillage(RT), were laid in a randomized complete block design. Concentrations of SOC and bulk density(BD) of the 0-80 cm soil layer were measured, and SOC stocks of the 0-20 and 0-80 cm soil layers were calculated on an equivalent soil mass(ESM) basis and fixed depth (FD) basis.Soil carbon budget(SCB) under diferent tillage systems were assessed on the basis of emissions of methane(CH4) and CO2 and the amount of carbon (C) removed by the rice harvest. After four years of experiment, the NT treatment sequestrated more SOC than the other treatments. The SOC stocks in the 0-80 cm layer under NT (on an ESM basis) was as high as 129.32 Mg C ha 1,significantly higher than those under PT and RT (P < 0.05). The order of SOC stocks in the 0-80 cm soil layer was NT > PT > RT,and the same order was observed for SCB; however, in the 0-20 cm soil layer, the RT treatment had a higher SOC stock than the PT treatment. Therefore, when comparing SOC stocks, only considering the top 20 cm of soil would lead to an incomplete evaluation for the tillage-induced efects on SOC stocks and SOC sequestrated in the subsoil layers should also be taken into consideration. The estimation of SOC stocks using the ESM instead of FD method would better reflect the actual changes in SOC stocks in the paddy filed, as the FD method amplified the tillage efects on SOC stocks. This study also indicated that NT plus straw retention on the soil surface was a viable option to increase SOC stocks in paddy soils.     

Metagenetics of fairy rings reveals complex and variable soil fungal communities

Although fairy rings are widely observed, little is known about the community processes associated with them. Here, we studied fairy rings in a natural grassland in northern Mongolia by sampling soils outside (future stage), directly under (present stage), and within (past stage) the rings, to represent different time stages during the expansion of fairy rings. Soil DNA was extracted for amplicon sequencing of the fungal ITS1 region. The present stage had reduced fungal diversity and different fungal communities toward the other stages. Most strikingly, there was an increase in the pathogenic fungus Gibberella baccata in the present stage of most rings and the saprotrophic fairy ring fungus Lepista luscina in several. However, no mushrooms of Lepista had been recorded from any of these rings during several years of observation. Known fairy ring fungi were not found in the present stage of every ring, even in some known rings to have previously displayed mushrooms of such species. It is possible that these fungi occurred or were more abundant at the unsampled leading edge of the ring. The increase in G. baccata is intriguing, but its importance, if any, is unclear. It is also unclear whether consortia of fungi or other microbes might be present in these rings. The absence or low abundance of the previously reported fairy ring fungal species suggests that their presence is transient, with rapid replacement by other fungi. No differences in soil parameters were found between the fairy ring stages, except for aluminum. There is a need for broader sampling, including analysis of non-fungal biota, to understand the functional diversity of fairy ring fungi and the consequences for plant communities.     

Effects of Long-Term Organic Amendments on Soil Organic Carbon in a Paddy Field A Case Study on Red Soil

Soil organic carbon （SOC） is one of the main carbon reservoirs in the terrestrial ecosystem. It is important to study SOC dynamics and effects of organic carbon amendments in paddy fields because of their vest expansion in south China. A study was carried out to evaluate the relationship between the SOC content and organic carbon input under various organic amendments at a long-term fertilization experiment that was established on a red soil under a double rice cropping system in 1981. The treatments included non-fertilization （CK）, nitrogen-phosphorus-potassium fertilization in early rice only （NPK）, green manure （Astragalus sinicus L.） in early rice only （OM1）, high rate of green manure in early rice only （OM2）, combined green manure in early rice and farmyard manure in late rice （OM3）, combined green manure in early rice, farmyard manure in late rice and rice straw mulching in winter （OM4）, combined green manure in early rice and rice straw mulching in winter （OMS）. Our data showed that the SOC content was the highest under OM3 and OM4, followed by OM1, OM2 and OM5, then NPK fertilization, and the lowest under non-fertilization. However, our analyses in SOC stock indicated a significant difference between OM3 （33.9 t ha^-1） and OM4 （31.8 t ha^-1）, but no difference between NPK fertilization （27 t ha^-1） and nonfertilization （28.1 t ha^-1）. There was a significant linear increase in SOC over time for all treatments, and the slop of linear equation was greater in organic manure treatments （0.276-0.344 g kg-1 yr^-1） than in chemical fertilizer （0.216 g kg^-1 yr^-1） and no fertilizer （0.127 g kg^-1 yr^-1）.     

Identiifcation and Molecular Mapping of the RsDmR Locus Conferring Resistance to Downy Mildew at Seedling Stage in Radish （Raphanus sativus L.）

Downy mildew （DM）, caused by the fungus Peronospora parasitica, is a destructive disease of radish （Raphanus sativus L.） worldwide. Host resistance has been considered as an attractive and environmentally friendly approach to control the disease. However, the genetic mechanisms of resistance in radish to the pathogen remain unknown. To determine the inheritance of resistance to DM, F1, F2 and BC1F1 populations derived from reciprocal crosses between a resistant line NAU-dhp08 and a susceptible line NAU-qtbjq-06 were evaluated for their responses to DM at seedling stage. All F1 hybrid plants showed high resistance to DM and maternal effect was not detected. The segregation for resistant to susceptible individuals statistically iftted a 3：1 ratio in two F2 populations （F2（SR） and F2（RS））, and 1：1 ratio in two BC1F1 populations, indicating that resistance to DM at seedling stage in radish was controlled by a single dominant locus designated as RsDmR. A total of 1 972 primer pairs （1 036 SRAP, 628 RAPD, 126 RGA, 110 EST-SSR and 72 ISSR） were screened, and 36 were polymorphic between the resistant and susceptible bulks, and consequently used for genotyping individuals in the F2 population. Three markers （Em9/ga24370, NAUISSR826700 and Me7/em10400） linked to the RsDmR locus within a 10.0 cM distance were identiifed using bulked segregant analysis （BSA）. The SRAP marker Em9/ga24370 was the most tightly linked one with a distance of 2.3 cM to RsDmR. These markers tightly linked to the RsDmR locus would facilitate marker-assisted selection and resistance gene pyramiding in radish breeding programs.     

Beyond Biochar-Soil,Plant and Environment: Research Progress and Future Prospects

Since industrial revolution, the atmospheric CO2 concentration has kept a continuous increase by more than 2.2 ppm yr^-1, and approaches to almost 400 ppm at present （Jouzel 2012）. China has become the largest country of greenhouse gas emission （GHG）, and confronts with great challenge to mitigate GHG.     

Determination of Residual Feed Intake and Its Associations with Single Nucleotide Polymorphism in Chickens

Marker assisted selection （MAS） for residual feed intake （RFI） is considered to be one of the powerful means to improve feed conversion efficiency, and therefore reduce production costs. To test the inner relationship among body compositions, growth traits and RFI, four models were proposed to assess the extensively explanatory variables accounting for partial variables in feed intake besides metabolic body weight and growth rate. As a result, the original model （Koch＇s model） had the lowest R2 （80.78%） and the highest Bayesian information criterion （1 323.3） value among the four models. Moreover, the effects on RFI caused by single nucleotide polymorphisms （SNPs） were assessed in this study. Twelve SNPs from 7 candidate genes were genotyped in 2 Chinese native strains, rs14743490 of RPLP2 gene showed suggestively significant association with initial body weight in both strains （P〈0.10）. rs15047274 of TAF15 was significantly associated with growth weight, final weight, and feed intake （P〈0.05） in N301 strain, in contrast, it was only suggestively significant associated with feed intake （P〈0.10） in N414 strain, rs15869967 was significantly associated with RFI in N414 strain but not in N301 strain. This study has identified potential genetic markers suitable for MAS in improving the above mentioned traits, but these associations need to be rectified in other larger populations in future.     

Genetic variation in progenies of 23 relic trees of Metasequoia glyptostroboides in their original habitat

Seeds of 23 relic Metasequoia glyptostroboides trees were randomly collected from their original, natural populations found in Lichuan City, Hubei Province, China. The seed characteristics and growth of their three-year-old progenies were investigated. Results show that the average intra-family coefficient of variation (CV) in height growth of the 23 three-year-old families was 14.53%, slightly higher than that of the inter-family CV (14.02%). Correspondingly, these two parameters for root-collar diameter growth were 15.44% and 13.15%. The 23 relic mother trees were significantly differed in their general combining abilities (GCA) and relative effect values of GCA (RGCA) of height growth and root-collar diameter growth over the three years. Both height and root-collar diameter growth had high family heritability in this seedling stage. After eliminating two inferior families, i.e., 15 and 46, 2.21% of genetic gain in height growth was obtained and 1.50% in root-collar diameter growth. It was also significantly different in crown size and number of branches among the 23 three-year-old seedling families. From a cluster analysis we concluded that eight families, i.e., 23, 28, 29, 30, 31, 33, 35 and 44 displayed superior GCA and RGCA in both height and root-collar diameter growth. Therefore, these relic mother trees displayed their potential as parents in seed orchards. With the exception of family 23, the other seven superior families were all collected at Guihua Village, Zhonglu Town, Lichuan City, which has been identified as an optimum district of M. glyptostroboides for seed collection.