Soybean cyst nematode (SCN, Heterodera glycines) is a devastating pathogen that infects soybean (Glycine max L. Merrill) and disrupts soybean production worldwide.  SCN infection upregulates or downregulates the expression of multiple genes in soybean.  However, the regulatory mechanisms that underlie these changes in gene expression remain largely unexplored.  N⁶-methyladenosine (m⁶A) methylation, one of the most prevalent mRNA modifications, contributes to transcriptional reprogramming during plant responses to pathogen infection.  Nevertheless, the role of m⁶A methylation in establishing compatible and incompatible soybean responses to SCN has not previously been studied.  Here, we performed transcriptome-wide m⁶A profiling of soybean roots infected with virulent and avirulent populations of SCN.  Compared with the compatible response, the incompatible response was associated with higher global m⁶A methylation levels, as well as more differentially modified m⁶A peaks (DMPs) and differentially expressed genes (DEGs).  A total of 133 and 194 genes showed significant differences in both transcriptional expression and m⁶A methylation levels in compatible and incompatible interactions; the most significantly enriched gene ontology terms associated with these genes were plant–pathogen interaction (compatible) and folate biosynthesis (incompatible).  Our findings demonstrate that the m⁶A methylation profiles of compatible and incompatible soybean responses are distinct and provide new insights into the regulatory mechanism underlying soybean response to SCN at the post-transcriptional modification level, which will be valuable for improving the SCN-resistant breeding.

Identification of the S genotype of Malus plants will greatly promote the discovery of new genes, the cultivation and production of apple, the breeding of new varieties, and the origin and evolution of self-incompatibility in Malus plants.  In this experiment, 88 Malus germplasm resources, such as Aihuahong, Xishuhaitang, and Reguanzi, were used as materials.  Seven gene-specific primer combinations were used in the genotype identification.  PCR amplification using leaf DNA produced a single S-RNase gene fragment in all materials.  The results revealed that 70 of the identified materials obtained a complete S-RNase genotype, while only one S-RNase gene was found in 18 of them.  Through homology comparison and analysis, 13 S-RNase genotypes were obtained: S₁S₂ (Aihuahong, etc.), S₁S₂₈ (Xixian Haitang, etc.), S₁S₅₁ (Hebei Pingdinghaitang), S₁S₃ (Xiangyangcun Daguo, etc.), S₂S₃ (Zhaiyehaitang, etc.), S₃S₅₁ (Xishan 1), S₃S₂₈ (Huangselihaerde, etc.), S₂S₂₈ (Honghaitang, etc.), S₄S₂₈ (Bo 11), S₇S₂₈ (Jiuquan Shaguo), S₁₀S_e (Dongchengguan 13), S₁₀S₂₁ (Dongxiangjiao) and S_eS₅₁ (Xiongyue Haitang).  Simultaneously, the frequency of the S gene in the tested materials was analyzed.  The findings revealed that different S genes had varying frequencies in Malus resources, as well as varying frequencies between intraspecific and interspecific.  S₃ had the highest frequency of 68.18%, followed by S₁ (42.04%).  In addition, the phylogenetic tree and origin evolution analysis revealed that the S gene differentiation was completed prior to the formation of various apple species, that cultivated species also evolved new S genes, and that the S₅₀ gene is the oldest S allele in Malus plants.  The S₁, S₂₉, and S₃₃ genes in apple-cultivated species, on the other hand, may have originated in M. sieversii, M. hupehensis, and M. kansuensis, respectively.  In addition to M. sieversii, M. kansuensis and M. sikkimensis may have also played a role in the origin and evolution of some Chinese apples.

The addition of silicon (Si) and organic fertilizers to soil conditioners can inhibit the transfer of heavy metal ions from soil to crops. However, it is not clear how Si and organic fertilizers affect soil properties and the micro-ecological environment and thereby reduce cadmium (Cd) accumulation in rice. In this study, the effects of L-type soil conditioners containing Si and organic fertilizers on bacterial and fungal community diversity, soil pH, organic matter, and available Si were analyzed with field experiments at two sites in Liuzhou City and Hezhou City, respectively, in Guangxi, China. With the increase of Si and organic fertilizer content in soil conditioner, rice yield respectively increased by 16.8–25.8 and 6.8–13.1%, and rice Cd content decreased significantly by 8.2–21.1 and 10.8–40.6%, respectively, at the two experimental sites. Soil microbiome analysis showed that the increase in abundance of Firmicutes and Actinobacteriota bacteria associated with Cd adsorption and sequestration, and Basidiomycota fungal populations associated with degradation of macromolecules favored the inhibition of soil Cd activity (soil exchangeable Cd decreased by 14.4–14.8 and 18.1–20.6%). This was associated with an increase in organic matter and Si content caused by applying soil conditioners. In conclusion, L-type soil conditioners, rich in Si and organic fertilizer, can reduce soil Cd bioavailability by regulating the dominant Cd passivating flora in the soil and ultimately reduce Cd accumulation in rice.

Soybean is one of the most important food crops worldwide.  Like other legumes, soybean can form symbiotic relationships with Rhizobium species.  Nitrogen fixation of soybean via its symbiosis with Rhizobium is pivotal for sustainable agriculture.  Type III effectors (T3Es) are essential regulators of the establishment of the symbiosis, and nodule number is a feature of nitrogen-affected nodulation.  However, genes encoding T3Es at quantitative trait loci (QTLs) related to nodulation have rarely been identified. Chromosome segment substitution lines (CSSLs) have a common genetic background but only a few loci with heterogeneous genetic information; thus, they are suitable materials for identifying candidate genes at a target locus.  In this study, a CSSL population was used to identify the QTLs related to nodule number in soybean.  Single nucleotide polymorphism (SNP) markers and candidate genes within the QTLs interval were detected, and it was determined which genes showed differential expression between isolines.  Four candidate genes (GmCDPK28, GmNAC1, GmbHLH, and GmERF5) linked to the SNPs were identified as being related to nodule traits and pivotal processes and pathways involved in symbiosis establishment.  A candidate gene (GmERF5) encoding a transcription factor that may interact directly with the T3E NopAA was identified.  The confirmed CSSLs with important segments and candidate genes identified in this study are valuable resources for further studies on the genetic network and T3Es involved in the signaling pathway that is essential for symbiosis establishment. 

With the increased cultivation of Bt crops in China, Apolygus lucorum and other mirid bugs have emerged as important agricultural pests because they are insensitive to the Bt proteins.  In addition, the reduction of pesticide applications after planting Bt crops also increases the severity of mirid bug outbreaks.  Peristenus spretus is a parasitoid of mirid nymphs, but its sensitivity to Bt proteins is not known.  In the current study, we developed a dietary exposure assay to assess the effects of Bt proteins (Cry1Ab, Cry1Ac, Cry1F, Cry2Aa, and Cry2Ab) on P. spretus adults using a diet consisting of a 10% honey solution with or without Bt proteins at 400 µg g^–1 diet.  The results showed that the survival and reproduction of P. spretus adults were reduced by the cysteine protease inhibitor E-64 (a positive control) but were not affected by any of the five Bt proteins.  The activities of digestive, detoxifying, and antioxidant enzymes in P. spretus were also unaffected by diets containing the Cry proteins, but they were significantly affected by the diet containing E-64.  We then developed a tri-trophic bioassay to determine the effects of the five Bt proteins on P. spretus larvae and pupae.  In this assay, A. lucorum nymphs fed an artificial diet containing Cry proteins were used as the hosts for P. spretus.  The results of the tri-trophic assay indicated that neither the pupation rate nor the eclosion rate of the P. spretus parasitoids were significantly affected by the presence of high concentrations of Bt proteins in the parasitized A. lucorum nymphs.  The overall results indicate that these two assays can be used to evaluate the toxicity of insecticidal compounds to P. spretus and that the tested Cry proteins are not toxic to P. spretus.  

Soybean (Glycine max (L.) Merr.), a typical short-day plant, is sensitive to photoperiod, which limits the geographical range for its cultivation.  In the flowering pathway regulated by photoperiod, E1, a flowering inhibitor in soybean, plays the dominant role in flowering time regulation.  Two E1 homologs, E1-like-a (E1La) and E1-like-b (E1Lb), play overlapping or redundant roles in conjunction with E1.  In the present study, E1 and E1La/b were simultaneously silenced via RNA interference (RNAi) in Zigongdongdou (ZGDD), an extremely late-flowering soybean landrace from southern China.  As a result, RNAi lines showed a much earlier-flowering phenotype and obvious photoperiod insensitivity compared with wild-type (WT) plants.  In RNAi transgenic plants, the expression levels of flowering inhibitor GmFT4 and flowering promoters GmFT2a/GmFT5a were significantly down- and up-regulated, respectively.  Further, the maturity group (MG) of the RNAi lines was reduced from WT ZGDD’s MG VIII (extremely late-maturity) to MG 000 (super-early maturity), which can even grow in the northernmost village of China located at a latitude of 53.5°N.  Our study confirms that E1 and E1La/b can negatively regulate flowering time in soybean.  The RNAi lines generated in this study, with early flowering and maturity traits, can serve as valuable materials and a technical foundation for breeding soybeans that are adapted to high-latitude short-season regions.

Red and blue light illumination has been reported to significantly affect plantlet growth.  Potato is an important food and feed crop in the world and potato plantlet cultured in vitro plays an important role in potato production.  However, few studies have documented the effects of red and blue light on the growth of potato plantlets revealed at the transcriptome level.  The objective of this study was to determine the growth and physiological responses of potato plantlets cultured in vitro under monochromatic red (RR), monochromatic blue (BB) as well as combined red and blue (RB) LEDs using the RNA-Seq technique.  In total, 3 150 and 814 differentially expressed genes (DEGs) were detected in potato plantlets under RR and BB, respectively, compared to RB (used as control).  Compared to the control, the DEGs enriched in “photosynthesis” and “photosynthesis-antenna proteins” metabolic pathways were up-regulated and down-regulated by BB and RR, respectively, which might be responsible for the increases and decreases of maximum quantum yield (F_v/F_m), photochemical quantum yield (φ_PSII), photochemical quenching (qP) and electron transfer rate (ETR) in BB and RR, respectively.  Potato plantlets exhibited dwarfed stems and extended leaves under BB, whereas elongated stems and small leaves were induced under RR.  These dramatically altered plantlet phenotypes were associated with variable levels of endogenous plant hormones gibberellin (GAs), indoleacetic acid (IAA) and cytokinins (CKs), as assessed in stems and leaves of potato plantlets.  In addition, monochromatic red and blue LEDs trigged the opposite expression profiles of DEGs identified in the “plant hormone signal transduction” metabolic pathway, which were closely related to the endogenous plant hormone levels in potato plantlets.  Our results provide insights into the responses of potato plantlets cultured in vitro to red and blue LEDs at the transcriptomic level and may contribute to improvements in the micro-propagation of potato plantlets cultured in vitro from the light spectrum aspect.

In this study, a safety enhanced Salmonella Pullorum (S. Pullorum) ghost was constructed using an antimicrobial peptide gene, and evaluated for its potential as a Pullorum disease (PD) vaccine candidate.  The antimicrobial peptide SMAP29 was co-expressed with lysis gene E to generate S. Pullorum ghosts.  No viable bacteria were detectable either in the fermentation culture after induction of gene E- and SMAP29-mediated lysis for 24 h or in the lyophilized ghost products.  Specific-pathogen-free (SPF) chicks were intraperitoneally immunized with ghosts at day 7 of age and no mortality, clinical symptoms or signs of PD such as anorexia, depression and diarrhea were observed.  On challenge with a virulent S. Pullorum strain at 4 wk post-immunization, a comparatively higher level of protection was observed in the S. Pullorum ghost immunized chickens with a minimum of pathological lesions and bacterial loads compared to the birds in inactivated vaccine groups.  In addition, immunization with the S. Pullorum ghosts induced a potent systemic IgG response and was associated with significantly increased levels of cytokine IFN-γ and IL-4 and relative percentages of CD4⁺ and CD8⁺ T lymphocytes.  Our results indicate that SMAP29 can be employed as a new secondary lethal protein to enhance the safety of bacterial ghosts, and to prepare a non-living bacterial vaccine candidate that can prevent PD in chickens.

Wheat crown rot caused by Fusarium spp. is a common disease worldwide.  Both Fusarium pseudograminearum and Fusarium graminearum infect wheat crown and produce mycotoxin leading to grain loss due to white head.  F. pseudograminearum (Fp) was reported in wheat from Henan Province of China a couple of years ago.  The wheat crown rot (CR) caused by this new pathogen is as an emerging severe disease of wheat, which has recently expanded to several provinces in China and is, therefore, under rapid investigation.  Colonization of wheat tissue by Fp is accomplished though the formation of a septated foot-shaped appressoria and generation of a penetration peg to break through the internal cells of leaf sheath.  The molecular mechanism by which Fp regulates the pathogenesis on wheat host is unclear.  Here, we report FpPDE1, a P-type ATPase-encoding predicted PDE1 orthologue gene of Magnaporthe oryzae, belonging to the DRS2 subfamily of aminophospholipid translocases.  The gene deletion of FpPDE1 with the split-marker approach did not obviously affect hyphae growth and conidiation, but led to an attenuated virulence on wheat base stem and root.  Our finding indicates that the putative aminophospholipid translocases is not essential for the infectious hyphae development in Fp.  

   In the rice cytoplasmic-genetic male sterility (CMS) system, the combination of a CMS line, maintainer line and restorer line carrying the restorer gene to restore fertility, is indispensable for the development of hybrids. However, the process of screening for the trait of fertility restoration is laborious and time-consuming. In the present study, we analyzed the nucleotide sequence of the Rf4 gene, which is the major locus controlling fertility restoration, to identify allele-specific variation. A single nucleotide polymorphism (SNP) A/C at +474 in the coding sequence (CDS) was found to be capable of strictly distinguishing groups of alleles Rf4 (A) and rf4 (C). Using KASP genotyping, this valuable SNP was converted to an allele-specific PCR marker. We evaluated and validated the marker among three-line parents with different backgrounds, and the results revealed a complete correlation between SNP alleles and the fertility restoration phenotype. Molecular screening was subsequently carried out for the presence of alleles of Rf4 and Rf3 among 328 diverse rice cultivars with worldwide distribution. The results demonstrate that this SNP marker could be the optimal choice for the molecular identification of potential restorers.

Plant structures and chemicals, which are developed from the shoot apical meristem (SAM), form the main barriers to insect feeding.  A plant chimera containing cells of different genetic origins in the SAM will be morphologically and chemically different compared with the parents and thus may result in differential resistance to herbivores.  In this study, we explore if particular elements of plant resistance are localized in one of the layers of SAM; the replacement of one cell layer in a chimera may be linked to change of a single resistance trait to herbivores.  The morphology and glucosinolate profiles of two periclinal chimeras (labeled as TTC and TCC, respectively) and grafted parents tuber mustard (labeled as TTT) and red cabbage (labeled as CCC) were compared and the performance of whitefly (Bemisia tabaci) in host selection, oviposition preference and development were assessed under controlled conditions.  Both chimeras possessed leaf trichomes as parent tuber mustard TTT, however, TTC had significantly more trichomes than TCC and parent TTT.  Leaf wax content of both chimeras was intermediate between the two parents.  Five aliphatic and two indole glucosinolates were detected in both chimeras, whereas three aliphatic glucosinolates (3-methyl-sulfinylpropyl, 4-methyl-sulfinylbutyl and 2-hydroxy-3-butenyl) were not detected in tuber mustard, and one aliphatic glucosinolate (3-butenyl) was not detected in red cabbage.  Unexpectedly for a chimera, the quantities of two aliphatic glucosinolates (3-methyl-sulfinylpropyl and 4-methyl-sulfinylbutyl) in both TTC and TCC were 3- to 5-fold higher than parents.  In olfactory preference assays, B. tabaci showed preference to CCC, followed by TCC, TTC and TTT, and number of eggs laid showed the same pattern: CCC>TCC>TTC>TTT.  Interestingly, more whiteflies landed on TTT plants than the other three types in a free choice experiment and the developmental duration from egg to adult was the shortest on TTT and increased in the order TTT<TTC<TCC<CCC.  Our results indicate plant defenses traits of leaf waxes, trichomes and glucosinolates are not controlled by one cell layer of SAM, but are influenced by interactions amongst cell layers.  The overall findings suggest that periclinal chimera systems can be a valuable approach for the study of plant-insect interactions and may also be useful for future resistance breeding. 

    Lead (Pb) contamination has often been recorded in Chinese field soils. In recent years, efforts have been made to investigate Pb toxicity thresholds in soils with plant growth and microbial assays. However, the influence of soil properties on Pb toxicity impacts on soil microbial processes is poorly understood. In this study ten soils with different properties were collected in China to investigate the relationships between thresholds of Pb toxicity to soil microbes and soil properties. The effect of soil leaching on Pb toxicity was also investigated to determine the possible influence of added anions on Pb toxicity during dose-response tests. Toxicity was inferred by measuring substrate-induced nitrification in leached and non-leached soils after Pb addition. We found that soil microbe Pb toxicity thresholds (EC_x, x=10, 50) differed significantly between the soils; the 10% inhibition ratio values (EC₁₀) ranged from 86 to 218 mg kg^–1 in non-leached soils and from 101 to 313 mg kg^–1 in leached soils. The 50% inhibition ratio values (EC₅₀) ranged from 403 to 969 mg kg^–1 in non-leached soils and from 494 to 1 603 mg kg^–1 in leached soils. Soil leaching increased EC10 and EC50 values by an average leaching factor (LF) of 1.46 and 1.33, respectively. Stepwise multiple regression models predicting Pb toxicity to soil microbes were developed based on EC_x and soil properties. Based on these models, soil pH and organic carbon are the most important soil properties affecting Pb toxicity thresholds (R₂>0.60). The quantitative relationship between Pb toxicity and soil properties will be helpful for developing soil-specific guidance on Pb toxicity thresholds in Chinese field soils.

   The variations of grain cadmiun (Cd) concentrations, translocation factors (TFs) of Cd from roots to shoots/grains of six rice cultivars, characterized with different Cd-sensitivities in polluted soil were studied, the selected rice cultivars were Xiangzao 17 (R1), Jiayu 211 (R2), Xiangzao 42 (R3), Zhuliangyou 312 (R4), Zhuliangyou 611 (R5), and Jinyou 463 (R6), respectively. The Cd subcellular distribution and Cd binding characteristics on subcellular fractions of rice root cell wall (CW) were further investigated. The results showed that the rice grain Cd contents varied significantly, with a maximum variation of 47.0% among the cultivars, the largest grain Cd content was observed with cultivar R1 (Cd-sensitivity cultivar) and the smallest with R5 (Cd-tolerance cultivar). The translocation factors of Cd from roots to shoots (TF_shoot) and roots to grains (TF_grain) varied greatly among the cultivars. In general, the TF_grain of the cultivars followed the order of R1>R2>R3>R4> R6-R5. The Cd concentration (mg kg^–1 FW) in the fraction of root CW, the fraction of cell wall removing pectin (CW-P) and the fraction of cell wall removing pectin and hemicellulose (CW-P-HC) of the cultivars generally followed the order of CW-P>CW>CW-P-HC; the ratios of Cd concentration (mg kg^–1 FW) in the fraction of CW-P to that of CW were mostly more than 1.10, while the ratios of Cd concentration in the fraction of CW-P-HC to that of CW were mostly less than 0.60, indicating that Cd was mainly stored in the hemicellulose of the root CW. The ratios of Cd of CW-P-HC to CW generally followed the descending order of R1~R2>R3>R4>R5~R6 for the cultivars, which implied that hemicellulose is probably the main subcellular pool for transferring Cd into rice grain, and it restrains the translocation of Cd from shoot to the grain, especially for the Cd-tolerance cultivars (R5 and R6), the compartmentation of more Cd in hemicellulose in root CW is probably one of the main mechanisms for Cd tolerance of rice cultivars.

It is imperative to derive an appropriate cadmium (Cd) health risk toxicity threshold for paddy soils to ensure the Cd concentration of rice grains meet the food safety standard. In this study, 20 rice cultivars from the main rice producing areas in China were selected, and a pot-experiment was conducted to investigate transformation of Cd in paddy soil-rice system with 0 (CK), 0.3 mg kg–1 (T1) and 0.6 mg kg–1 (T2) Cd treatments in greenhouse. The results showed that Cd concentrations of rice grains existed significant difference (P<0.05) in 20 rice cultivars under the same Cd level in soil. The Cd concentrations of rice grains of the CK, T1 and T2 treatments were in the range of 0.143–0.202, 0.128–0.458 and 0.332–0.806 mg kg–1, respectively. Marked differences of the ratios of Cd concentration for soil to rice grain (BCFs) and transfer factors (TFs, root to grain and straw to grain) among the tested cultivars were observed in this study. The bioconcentration factors (BCFgrain) and TFs of the 20 rice cultivars were 0.300–1.112 and 0.342–0.817, respectively. The TFs of Cd from straw to grain ranged from 0.366 to 1.71, with significant differences among these 20 rice cultivars. The bioconcentration factors (BCFgrain) and TFs among the 20 rice cultivars ranged from 0.300–1.112 and 0.342–0.817, respectively. The species-sensitivity distribution (SSD) of Cd sensitivity of the rice species could be fitted well with Burr-III (R2=0.987) based on the data of BCFs. The toxicity threshold of Cd derived from SSD for the paddy soil was 0.507 mg kg–1 in the present study.

Rapeseed (Brassica napus L.) oil is the crucial source of edible oil in China. In addition, it can become a major renewable and sustainable feedstock for biodiesel production in the future. It is known that photosynthesis products are the primary sources for dry matter accumulation in rapeseed. Therefore, increasing the photosynthetic efficiency is desirable for the raise of rapeseed yield. The objective of the present study was to identify the genetic mechanism of photosynthesis based on the description of relationships between different photosynthetic traits and their quantitative trait loci (QTL) by using a recombinant inbred line (RIL) population with 172 lines. Specifically, correlation analysis in this study showed that internal CO2 concentration has negative correlations with other three physiological traits under two different stages. Totally, 11 and 12 QTLs of the four physiological traits measured at the stages 1 and 2 were detected by using a high-density single nucleotidepolymorphism (SNP) markers linkage map with composite interval mapping (CIM), respectively. Three co-localized QTLs on A03 were detected at stage 1 with 5, 5, and 10% of the phenotypic variation, respectively. Other two co-localized QTLs were located on A05 at stage 2, which explained up to 12 and 5% of the phenotypic variation, respectively. The results are beneficial for our understanding of genetic control of photosynthetic physiological characterizations and improvement of rapeseed yield in the future.

In order to reveal the impact of various fertilization strategies on carbon (C) and nitrogen (N) accumulation and allocation in corn (Zea mays L.), corn was grown in the fields where continuous fertilization management had been lasted about 18 years at two sites located in Central and Northeast China (Zhengzhou and Gongzhuling), and biomass C and N contents in different organs of corn at harvest were analyzed. The fertilization treatments included non-fertilizer (control), chemical fertilizers of either nitrogen (N), or nitrogen and phosphorus (NP), or phosphorus and potassium (PK), or nitrogen, phosphorus and potassium (NPK), NPK plus manure (NPKM), 150% of the NPKM (1.5NPKM), and NPK plus straw (NPKS). The results showed that accumulated C in aboveground ranged from 2 550–5 630 kg ha–1 in the control treatment to 9 300–9 610 kg ha–1 in the NPKM treatment, of which 57–67% and 43–50% were allocated in the non-grain organs, respectively. Accumulated N in aboveground ranged from 44.8–55.2 kg ha–1 in the control treatment to 211–222 kg ha–1 in the NPKM treatment, of which 35–48% and 33–44% were allocated in the non-grain parts, respectively. C allocated to stem and leaf for the PK treatment was 65 and 49% higher than that for the NPKM treatment at the both sites, respectively, while N allocated to the organs for the PK treatment was 18 and 6% higher than that for the NPKM treatment, respectively. This study demonstrated that responses of C and N allocation in corn to fertilization strategies were different, and C allocation was more sensitive to fertilization treatments than N allocation in the area.

The Bacillus thuringiensis vegetative insecticidal protein, Vip3A, represents a new family of Bt toxin and is currently applied to commercial transgenic cotton. To determine whether the Cry1Ac-resistant Helicoverpa armigera is cross-resistant to Vip3Aa protein, insecticidal activities, proteolytic activations and binding properties of Vip3Aa toxin were investigated using Cry1Ac-susceptible (96S) and Cry1Ac-resistant H. armigera strain (Cry1Ac-R). The toxicity of Vip3Aa in Cry1Ac-R slightly reduced compared with 96S, the resistance ratio was only 1.7-fold. The digestion rate of full-length Vip3Aa by gut juice extracts from 96S was little faster than that from Cry1Ac-R. Surface plasmon resonance (SPR) showed there was no significant difference between the binding affinity of Vip3Aa and BBMVs between 96S and Cry1Ac-R strains, and there was no significant competitive binding between Vip3Aa and Cry1Ac in susceptible or resistant strains. So there had little cross-resistance between Vip3Aa and Cry1Ac,Vip3A+Cry proteins maybe the suitable pyramid strategy to control H. armigera in China in the future.

Two experiments were conducted to determine the effects of fiber level from alfalfa meal and sampling time on the apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of amino acids (AA) in growing pigs. A total of 24 ileal-cannulated pigs (Duroc×(Large White×Landrace) with body weight (21.4±1.5) kg) were randomly allotted to 4 treatments. The pigs were provided a corn-soybean meal diet or a diet containing 5, 10 or 20% of alfalfa meal during two 10-d experimental periods. The AID of AA was measured. Six ileal-cannulated pigs were fed a protein-free diet in order to estimate the endogenous protein losses and SID of AA. Ileal AA digestibility was not affected by inclusion of 5 or 10% alfalfa meal in the diet (P>0.05). The AID of His, Lys, Met, Phe, Thr, Val, Ala, Asp, Cys, Gly, Pro, Ser and Tyr reduced by 2.0–6.8% with the addition level of alfalfa meal (linear, P<0.05). The SID of His, Lys, Ser, Thr and Tyr decreased by 2.2–4.3% as the level of alfalfa meal in the diet increased (linear, P<0.05). The AID and SID of AA were not affected by the sampling time (P>0.05). A multiple linear regression analysis, taking into account both the soluble and insoluble fiber content in the diets, explained more than 36% of variation in SID of Ser and Thr (P<0.05). In conclusion, the corn-soybean meal diet containing 10% of alfalfa meal did not affect ileal AA digestion. The AID and SID of AA were similar between two sampling times. Increasing the concentration of total dietary fiber from 12.3 to 21.4% by adding graded levels of alfalfa meal (0–20%) to a corn-soybean meal control diet induced a linear reduction in AID and SID of most AA. Soluble and insoluble fibers from alfalfa meal have differential roles in the AA digestion, which may help explain the variation observed in the SID of partial AA. These findings would provide important information for dietary fiber level and composition related to AA digestion.

Seeds play a central role in the life cycle of plants. Seed hardness in pomegranates is of economic relevance, yet scarcely studied and poorly understood in China. In this study, we compared the proteomic differences between Zhongnonghong (soft-seeded) and Sanbai (hard-seeded) pomegranates. A total of 892 protein spots from both varieties were detected on two-dimensional electrophoresis gels (2-DE); 76 spots showed greater than a 1.5-fold or less than a 0.66-fold difference (P<0.05) in Zhongnonghong compared to Sanbai, of which 24 exhibited greater than a 2-fold change. Compared with Sanbai, Zhongnonghong possessed 14 up-regulated, and 10 down-regulated proteins. We identified and annotated 5 of these by using MALDI-TOF-TOF MS: pyruvate dehydrogenase (PDH) E1-β family protein (spot 4 609); alanine aminotransferase 2-like (ALT2L); mitochondrial glycine decarboxylase complex P-protein (spot 5 803); phosphofructokinase B (PfkB)-type family of carbohydrate kinase (spot 8 411); and putative dnaK-type molecular chaperone heat shock cognate protein 70 (Hsc70) (spot 9 006). Of these, 3 proteins (spots 4 609, 5 608, 5 803) were hypothesized to play a role in the formation of seed hardness. The other two proteins (spots 8 411, 9 006) were theorized to play a role in protecting the seeds from adverse stress during periods of fruit maturation. This study sets the foundation for further research on molecular mechanisms related to pomegranate seed hardness.

The improvement of soil productivity depends on a rational input of water and nutrients, optimal field management, and the increase of basic soil productivity (BSP). In this study, BSP is defined as the productive capacity of a farmland soil with its own physical and chemical properties for a specific crop season under local field management. Based on 19-yr data of the long-term agronomic experiments (1989–2008) on a fluvo-aquic soil in Zhengzhou, Henan Province, China, the decision support system for agrotechnology transfer (DSSAT ver. 4.0) crop growth model was used to simulate yields by BSP of winter wheat (Triticum aestivium L.) and summer maize (Zea mays L.) to examine the relationship between BSP and soil organic carbon (SOC) under long-term fertilization. Five treatments were included: (1) no fertilization (control), (2) nitrogen, phosphorus and potassium fertilizers (NPK), (3) NPK plus manure (NPKM), (4) 1.5 times of NPKM (1.5NPKM), and (5) NPK plus straw (NPKS). After 19 yr of treatments, the SOC stock increased 16.7, 44.2, 69.9, and 25.2% under the NPK, NPKM, 1.5NPKM, and NPKS, respectively, compared to the initial value. Among various nutrient factors affecting contribution percentage of BSP to winter wheat and summer maize, SOC was a major affecting factor for BSP in the fluvo-aquic soil. There were significant positive correlations between SOC stock and yields by BSP of winter wheat and summer maize (P<0.01), and yields by BSP of winter wheat and summer maize increased 154 and 132 kg ha–1 when SOC stock increased 1 t C ha–1. Thus, increased SOC accumulation is a crucial way for increasing BSP in fluvo-aquic soil. The manure or straw combined application with chemical fertilizers significantly enhanced BSP compared to the application of chemical fertilizers alone.

The fiber level and composition have an important effect on nutrient digestibility of swine diets. Little information is known about the effects of fiber level and composition from alfalfa meal on nutrient digestibility of fattening pigs fed a corn-soybean meal-based diet. The objective of this experiment was to determine the effects of alfalfa fiber on the growth performance, intestinal nutrient flow and apparent total tract digestibility (ATTD) of nutrients in fattening pigs. 24 barrows (Duroc×(Large White×Landrace), body weight=(60.6±0.7) kg) were randomly allotted to 4 treatments with 6 replicates of 1 pig per replicate. The pigs were provided a control diet or a diet containing 5, 10 or 20% of alfalfa meal during a 14-d experiment period. Average daily gain (ADG) and the ATTD of dry matter (DM), organic matter (OM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF) and gross energy (GE) reduced linearly as the level of alfalfa meal in the diet increased (P<0.01). The total tract flow of DM, OM, CP, NDF, ADF, and GE increased with the increase in dietary alfalfa (linear, P<0.05). Growth performance and nutrient digestion were not affected by inclusion of 5% alfalfa meal in the diet (P>0.05). A multiple linear regression analysis, taking into account both soluble and insoluble fiber intake, explained approximately 70% of the variation in the ATTD of DM, OM, NDF, and GE (P<0.01). In conclusion, alfalfa meal should be limited to less than 5% of the diet in fattening pigs to maximize growth performance and nutrient digestion. Soluble and insoluble fiber from alfalfa meal has the differential roles in nutrient digestion, which may help explain the main variation observed in nutrient digestibility. These findings suggest that knowledge of specific fiber components is necessary to accurately predict the effects of dietary fiber on nutrient digestibility.

Nitric oxide (NO) is a gaseous signaling molecule in plants that plays a key role in mediating a wide range of physiological processes and responses to biotic and abiotic stresses. The present study was conducted to investigate the effects of the exogenous application of sodium nitroprusside (SNP), an NO donor, on cadmium (Cd)-induced oxidative stress and Cd uptake in rice plants. Rice plants were exposed to Cd stress (0.2 mmol L-1 CdCl2) and different concentrations of SNP (0.05, 0.1, 0.2, and 0.4 mmol L-1). A SNP concentration of 0.1 mmol L-1 (SNP10) significantly reduced the Cd-induced decrease in shoot and root dry weights and leaf chlorophyll concentrations. The addition of NO also reduced the malondialdehyde (MDA), hydrogen peroxide (H2O2) and ascorbic acid (ASA) concentrations. However, the reduction in glutathione (GSH) concentration was inhibited by NO treatment. Moreover, NO prevented the Cd-induced increase in antioxidative enzyme activity. The amount of Cd accumulation in rice plants was also influenced by the addition of NO. The NO supplied by the SNP enhanced the Cd tolerance of the rice by increasing the Cd uptake by the roots and decreasing the Cd accumulation by the shoots. However, the application of potassium ferrocyanide (Cd+Fe) or sodium nitrate and nitrite (Cd+N) (without NO release), did not exhibit the effects of the SNP. Furthermore, the effects of the SNP were reversed by the addition of hemoglobin (an NO scavenger). Our results suggested that exogenous NO was involved in the resistance of rice to Cdtoxicity.

Kunming mouse strain is widely used in China, and the superovulation was administrated with 10 IU PMSG combined with 10 IU hCG. In this study, the effects of the exogenous gonadotropins on superovulation of Kunming mice and embryo quality derived from the superovulated mice were assessed. Female mice at 6-8-wk old were superovulated with 0, 5, 7.5 and 10 IU PMSG/hCG and mated with male mice. The embryos were retrieved at 2.5 d post coitum. No statistic difference was observed for the number of 2-cell embryos collected per mouse between control and 5 IU PMSG/hCG treatment group, but the number significantly increased for 7.5 and 10 IU PMSG/hCG treatment group (P<0.05). The average number of 4- cell and 8-cell embryos collected from each mouse significantly differed between control and 5, 7.5, 10 IU PMSG/hCG treatment groups (P<0.05). When 8-cell embryos derived from mice administrated with 0, 5, 7.5 and 10 IU PMSG/hCG were cultured in KSOM, the blastocyst development rates were 88.1, 94.7, 96.1 and 94.3%, respectively, which were similar to control (P>0.05). This indicated that exogenous gonadotropins have no effects on development of Kunming mouse embryos. The quality of blastocyst was assessed by labelling with Hoechst and propidium iodide for inner cell mass and trophectoderm cells, the result showed that ICM/TE ratio significantly decreased for 10 IU PMSG/hCG treatment group compared with control, 5 and 7.5 IU PMSG/hCG treatment group (P<0.05). This suggested that the embryo quality of Kunming mouse has been affected by high dose of gonadotropins.