Salinity tolerance is an important physiological index for crop breeding.  Roots are typically the first plant tissue to withstand salt stress.  In this study, we found that the tomato (Solanum lycopersicum) trehalose-6-phosphate phosphatase (SlTPP4) gene is induced by abscisic acid (ABA) and salt, and is mainly expressed in roots.  Overexpression of SlTPP4 in tomato enhanced tolerance to salt stress, resulting in better growth performance.  Under saline conditions, SlTPP4 overexpression plants demonstrated enhanced sucrose metabolism, as well as increased expression of genes related to salt tolerance.  At the same time, expression of genes related to ABA biosynthesis and signal transduction was enhanced or altered, respectively.  In-depth exploration demonstrated that SlTPP4 enhances Casparian band development in roots to restrict the intake of Na⁺.  Our study thus clarifies the mechanism of SlTPP4-mediated salt tolerance, which will be of great importance for the breeding of salt-tolerant tomato crops.

Peanut pod shape is a heritable trait which affects the market acceptance of in-shell peanut products.  In order to determine the genetic control of pod shape, six component traits of pod shape (pod length, pod width, pod length/width ratio, pod roundness, beak degree and constriction degree) were measured using an image-based phenotyping method.  A recombinant inbred line (RIL) population consisting of 181 lines was phenotyped across three environments.  Continuous distributions and transgressive segregations were demonstrated in all measured traits and environments.   Significant correlations were found among most component traits with broad-sense heritability ranging from 0.87 to 0.95.  Quantitative trait locus (QTL) analysis yielded 26 additive QTLs explaining 3.79 to 52.37% phenotypic variations.  A novel, stable and major QTL region conditioning multiple shape features was detected on chromosome 2, which spans a 10.81-Mb genomic region with 543 putative genes.  Bioinformatics analysis revealed several candidate genes in this region.  In addition, 73 pairs of epistatic interactions involving 92 loci were identified for six component traits explaining 0.94–6.45% phenotypic variations.  These results provide new genetic loci to facilitate genomics-assisted breeding of peanut pod shape.

Genetic improvement has promoted wheat’s grain yield and nitrogen use efficiency (NUE) during the past decades. Therefore, the current wheat cultivars exhibit higher grain yield and NUE than previous cultivars in the Yangtze River Basin, China since the 2000s. However, the critical traits and mechanisms of the increased grain yield and NUE remain unknown. This study explores the mechanisms underlying these new cultivars’ increased grain yield and NUE by studying 21 local cultivars cultivated for three growing seasons from 2016 to 2019. Significantly positive correlations were observed between grain yield and NUE in the three years. The cultivars were grouped into high (HH), medium (MM), and low (LL) grain yield and NUE groups. The HH group exhibited significantly high grain yield and NUE. High grain yield was attributed to more effective ears by high tiller fertility and greater single-spike yield by increasing postanthesis single-stem biomass. Compared to other groups, the HH group demonstrated a longer leaf stay-green ability and a greater flag leaf photosynthetic rate after anthesis. It also showed higher N accumulation at pre-anthesis, which contributed to increasing N accumulation per stem, including stem and leaf sheath, leaf blade, and unit leaf area at preanthesis, and promoting N uptake efficiency, the main contribution of high NUE. Moreover, tiller fertility was positively related to N accumulation per stem, N accumulation per unit leaf area, leaf stay-green ability, and flag leaf photosynthetic rate, which indicates that improving tiller fertility promoted N uptake, leaf N accumulation, and photosynthetic ability, thereby achieving synchronous improvements in grain yield and NUE. Therefore, tiller fertility is proposed as an important kernel indicator that can be used in the breeding and management of cultivars to improve agricultural efficiency and sustainability.

This paper aims to optimize the culling compensation policy from a micro perspective through scenario simulation.  Based on an investigation of 273 pig farms in eight regions, four typical pig farms were constructed according to farm size and breeding mode, representing the swine producers in China.  Besides, a decision objective function of pig farms facing suspected African swine fever (ASF) outbreaks was constructed.  This study used a mathematical programming model to design and simulate scenarios based on compensation standards and local implementation levels, aiming to incentivize pig farms to report epidemics.  The results show that the optimal decisions on epidemic reports differed among typical farms and by herd daily age.  The results suggest the following adjustments for optimizing culling compensation policies: (1) to set culling compensation standards based on the market value and (2) to maintain a high level of epidemic surveillance capability in the animal husbandry and veterinary sector.

Soybean yield has been increased through high planting density, but investigating plant height and petiole traits to select for compact architecture, lodging resistance, and high yield varieties is an underexplored avenue to improve yield.  We compared the relationship between yield-related traits, lodging resistance, and petiole-associated phenotypes in the short petiole germplasm M657 with three control accessions over 2017-2018 in four locations of the Huang-Huai region.  The results showed M657 exhibited stable and high tolerance to high planting density and resistance to lodging, especially at the highest density (8×10⁵ plants ha^-1).  Regression analysis showed that shorter petiole length was significantly associated with increased lodging resistance.  Yield analysis showed that M657 achieved higher yields under higher densities, especially in the north Huang-Huai region.  There are markedly different responses to intra- and inter-row spacing designs among varieties in both lodging and yield related to location and density.  Lodging was positively correlated with planting density, plant height, petiole length, and number of effective branches, and negatively correlated with stem diameter, seed number per plant, and seed weight per plant.  The yield of soybean was increased by appropriately increasing planting density on the basis of current soybean varieties in the Huang-Huai region.  This study provides a valuable new germplasm resource for introgression of compact architecture traits amenable to high yield in high density planting systems and establishes a high-yield model of soybean in the Huang-Huai region.

The B-box (BBX) family of proteins consists of zinc-finger transcription factors with one or two highly conserved B-box motifs at their N-termini.  BBX proteins play crucial roles in various aspects of plant growth and development, including seedling photomorphogenesis, shade avoidance, flowering time, and biotic and abiotic stress responses.  Previous studies have identified many different BBXs from several plant species, although the BBX family members in maize are largely unknown.  Genome-wide identification and comprehensive analysis of maize BBX (ZmBBX) expression and interaction networks would therefore provide valuable information for understanding their functions.  In this study, 36 maize BBXs in three major clades were identified.  The ZmBBXs within a given clade were found to share similar domains, motifs, and genomic structures.  Gene duplication analyses revealed that the expansion of BBX proteins in maize has mainly occurred by segmental duplication.  The expression levels of ZmBBXs were analyzed in various organs and tissues, and under different abiotic stress conditions.  Protein–protein interaction networks of ZmBBXs were established using bioinformatic tools and verified by bimolecular fluorescence complementation (BiFC) assays.  Our findings can facilitate a greater understanding of the complexity of the ZmBBX family and provide novel clues for unravelling ZmBBX protein functions

Phenotypic screening of soybean germplasm suitable for high planting density is currently the most viable strategy to increase yield.  Previous studies have shown that soybean varieties with dwarf features and a short petiole often exhibit a compact plant architecture which could improve yield through increased planting density, although previously reported short petiole accessions were ultimately not usable for breeding in practice.  Here, we established a method to assess petiole length and identified an elite mutant line, M657, that exhibits high photosynthetic efficiency.  The agronomic traits of M657 were evaluated under field conditions, and appeared to be stable for short petiole across seven locations in northern, Huang–Huai, and southern China from 2017 to 2018.  Compared with the Jihuang 13 wild type, the mutant M657 was shorter in both petiole length and plant height, exhibited lower total area of leaf, seed weight per plant and 100-seed weight, but had an increased number of effective branches and the growth period was prolonged by 2–7 days.  Using M657 as a parental line for crosses with four other elite lines, we obtained four lines with desirable plant architecture and yield traits, thus demonstrating the feasibility of adopting M657 in breeding programs for soybean cultivars of high density and high yield.

Crop planting patterns are an important component of agricultural land systems.  These patterns have been significantly changed due to the combined impacts of climatic changes and socioeconomic developments.  However, the extent of these changes and their possible impacts on the environment, terrestrial landscapes and rural livelihoods are largely unknown due to the lack of spatially explicit datasets including crop planting patterns.  To fill this gap, this study proposes a new method for spatializing statistical data to generate multitemporal crop planting pattern datasets.  This method features a two-level model that combines a land-use simulation and a crop pattern simulation.  The output of the first level is the spatial distribution of the cropland, which is then used as the input for the second level, which allocates crop censuses to individual gridded cells according to certain rules.  The method was tested using data from 2000 to 2019 from Heilongjiang Province, China, and was validated using remote sensing images.  The results show that this method has high accuracy for crop area spatialization.  Spatial crop pattern datasets over a given time period can be important supplementary information for remote sensing and thus support a wide range of application in agricultural land systems.

The potato cyst nematodes (PCN) Globodera rostochiensis (Wollenweber) Skarbilovich, 1959 is considered the most damaging nematode pest of potato worldwide that causes significant yield losses, and this nematode is recognized and listed as a quarantine nematode in many countries (EPPO 2017).  China is currently the largest producer of potato in the world, while the total production is also the highest (Guan and Cai 2019).  The survey for cyst nematodes on potato were conducted in Yunnan and Sichuan provinces of China during 2018–2020, numerous cysts were observed on potato roots in Huize County and Ludian County of Yunnan Province, Zhaojue County and Yuexi County of Sichuan Province.  Cysts and second-stage juveniles (J2s) were isolated from each soil sample using the Cobb decanting and sieving method.  The morphology of cysts and J2s and molecular analysis established the identity of this species as golden cyst nematode Globodera rostochiensis (Subbotin et al. 2010).  For morphological analysis, the cysts were characterized by smoothly rounded with a small projecting neck, brown and golden color, terminal cone was absent and circumfenestrate.  The key morphometrics of cysts (n=25) were: length excluding neck 705±24 (689–747) μm, width 698±28 (678–759) μm, number of cuticular ridges between anus and vulval fenestra 17.3±1.7 (14–19); fenestral diameter 13.6±1.1 (12.25–15.45) μm; distance from anus to the edge of fenestra 63.7±11.3 (48.23–79.14) μm; Granek’s ratio 4.7±0.7 (3.92–5.75).  The key morphometrics of J2s (n=25): body length 453.9±16.6 (440–496) μm, stylet length 21.9±1.0 (20.3–24.3) μm, tail length 51.1±3.2 (45.5–55.5) μm, and hyaline region length 24.4±2.5 (21.7–29.9) μm.  Morphology of the cysts and J2 were consistent with those of G. rostochiensis (Subbotin et al. 2010; EPPO 2017).  Moreover, the identification result was confirmed by PCR using universal primers TW81 (5´-GTTTCCGTAGGTGAACCTGC-3´) and AB28 (5´-ATATGCTTAAGTTCAGCGGGT-3´) for ITS region and D2A (5´-TTTTTTGGGCATCCTGAGGTTTAT-3´) D3B (5´-AGCACCTAAACTTAAAACATAATGAAAATG-3´) for rDNA-28S region, respectively.  The ITS rDNA sequences (GenBank accessions MZ042365, MZ042366, MZ042369, and MZ042370) exhibited 99.83% identity match to G. rostochiensis sequences available in the GenBank (GQ294513).  Sequence from the 28S region (GenBank accessions MZ057595, MZ057596, MZ057599, and MZ057600) was 99.33% similar to those of G. rostochiensis isolate from MF773722.  The species was also confirmed with species-specific primers ITS5 (5´-GGAAGTAAAAGTCGTAACAAGG-3´) and PITSr3 (5´-AGCGCAGACATGCCGCAA-3´) (Bulman and Marshall 1997), a single 434-bp fragment was obtained from Huize, Ludian, Zhaojue and Yuexi populations.  The pathog enicity testing of Huize, Ludian, Zhaojue and Yuexi, three weeks-old potato plants (cv. Qinshu 9)

were inoculated with 2 000 eggs, and cultured in an incubator at 23°C/20°C with a 16 h/8 h light/dark photoperiod.  After three months inoculation, 36±7.2 cysts and females were extracted from the infested potato roots, no females and cysts were observed on control plants.  

This is the first report of potato golden cyst nematode G. rostochiensis in China.  

Cell wall architecture plays a key role in stalk strength and forage digestibility.  Lignin, cellulose, and hemicellulose are the three main components of plant cell walls, and they can impact stalk quality by affecting the structure and strength of the cell wall.  To explore cell wall development during secondary cell wall lignification in maize stalks, conventional and conditional genetic mapping were used to identify the dynamic quantitative trait loci (QTLs) of the cell wall components and digestibility traits during five growth stages after silking.  Acid detergent lignin (ADL), cellulose (CEL), acid detergent fiber (ADF), neutral detergent fiber (NDF), and in vitro dry matter digestibility (IVDMD) were evaluated in a maize recombinant inbred line (RIL) population.  ADL, CEL, ADF, and NDF gradually increased from 10 to 40 days after silking (DAS), and then they decreased.  IVDMD initially decreased until 40 DAS, and then it increased slightly.  Seventy-two QTLs were identified for the five traits, and each accounted for 3.48–24.04% of the phenotypic variation.  Six QTL hotspots were found, and they were localized in the 1.08, 2.04, 2.07, 7.03, 8.05, and 9.03 bins of the maize genome.  Within the interval of the pleiotropic QTL identified in bin 1.08 of the maize genome, six genes associated with cell wall component biosynthesis were identified as potential candidate genes for stalk strength as well as cell wall-related traits.  In addition, 26 conditional QTLs were detected in the five stages for all of the investigated traits.  Twenty-two of the 26 conditional QTLs were found at 30 DAS conditioned using the values of 20 DAS, and at 50 DAS conditioned using the values of 40 DAS.  These results indicated that cell wall-related traits are regulated by many genes, which are specifically expressed at different stages after silking.  Simultaneous improvements in both forage digestibility and lodging resistance could be achieved by pyramiding multiple beneficial QTL alleles identified in this study.

Meloidogyne vitis is a new root-knot nematode parasitic on grape root in Yunnan Province, China.  In order to establish a rapid, reliable and specific molecular detection method for M. vitis, the species-specific primers were designed with rDNA-ITS (ribosomal DNA internal transcribed spacer) gene fragment as the target.  The reaction system was optimized and the reliability, specificity and sensitivity of primer were testified, therefore, a rapid PCR detection method for M. vitis was established.  The result showed that the optimal annealing temperature of the primers was 53°C, which was suitable for the detection of different life stages of M. vitis.  Specificity test showed that the specific fragment size of 174 bp was obtained from M. vitis, but other five non-target nematodes did not have any amplification bands, thus effectively distinguish M. vitis and the other five species, and could specifically detect the M. vitis from mixed populations.  Sensitivity test showed that this PCR technique could detect the DNA of a single second-stage juvenile (J₂) and 10^–4 female.  Futhermore, this PCR technique could be used to detect directly M. vitis from soil samples.  The rapid, sensitive and specific PCR molecular detection technique could be used for the direct identification of a single J₂ of M. vitis and the detection of M. vitis in mixed nematode populations and the detection of two J₂s or one male in 0.5 g soil samples, which will provide technical support for the investigation of the occurrence and damage of M. vitis and the formulation of efficient green control strategies.

Understanding the effects of land use/cover change (LUCC) on regional climate is critical for achieving land use system sustainability and global climate change mitigation.  However, the quantitative analysis of the contribution of LUCC to the changes of climatic factors, such as precipitation & temperature (P&T), is lacking.  In this study, we combined statistical methods and the gravity center model simulation to quantify the effects of long-term LUCC on P&T in the Songnen Plain (SNP) of Northeast China from 1980–2018.  The results showed the spatiotemporal variability of LUCC. For example, paddy field had the largest increase (15 166.43 km²) in the SNP, followed by dry land, while wetland had the largest decrease (19 977.13 km²) due to the excessive agricultural utilization and development.  Annual average precipitation decreased at a rate of –9.89 mm per decade, and the warming trends were statistically significant with an increasing rate of 0.256°C per decade in this region since 1980.  The model simulation revealed that paddy field, forestland, and wetland had positive effects on precipitation, which caused their gravity centers to migrate towards the same direction accompanied by the center of precipitation gravity, while different responses were seen for building land, dry land and unused land.  These results indicated that forestland had the largest influence on the increase of precipitation compared with the other land use types.  The responses in promoting the temperature increase differed significantly, being the highest in building land, and the lowest in forestland.  In general, the analysis of regional-scale LUCC showed a significant reduction of wetland, and the increases in building land and cropland contributed to a continuous drying and rapid warming in the SNP.

Improving both grain yield and resource use efficiencies simultaneously is a major challenge in rice production. However, few studies have focused on integrating dense planting with delayed and reduced nitrogen application to enhance grain yield, nitrogen use efficiency (NUE) and radiation use efficiency (RUE) in rice (Oryza sativa L.) in the double rice cropping system in South China. A high-yielding indica hybrid rice cultivar (Yliangyou 143) was grown in field experiments in Guangxi, South China, with three cultivation managements: farmers’ practice (FP), dense planting with equal N input and delayed N application (DPEN) and dense planting with reduced N input and delayed N application (DPRN). The grain yields of DPRN reached 10.6 and 9.78 t ha^–1 in the early and late cropping seasons, respectively, which were significantly higher than the corresponding yields of FP by 23.9–29.9%. The grain yields in DPEN and DPRN were comparable. NUE in DPRN reached 65.2–72.9 kg kg^–1, which was 61.2–74.1% higher than that in FP and 24.6–30.2% higher than that in DPEN. RUE in DPRN achieved 1.60–1.80 g MJ^–1, which was 28.6–37.9% higher than that in FP. The productive tiller percentage in DPRN was 7.9–36.2% higher than that in DPEN. Increases in crop growth rate, leaf area duration, N uptake from panicle initiation to heading and enhancement of the apparent transformation ratio of dry weight from stems and leaf sheaths to panicles all contributed to higher grain yield and higher resource use efficiencies in DPRN. Correlation analysis revealed that the agronomic and physiological traits mentioned above were significantly and positively correlated with grain yield. Comparison trials carried out in Guangdong in 2018 and 2019 also showed that DPRN performed better than DPEN. We conclude that DPRN is a feasible approach for simultaneously increasing grain yield, NUE and RUE in the double rice cropping system in South China.

Rice, wheat and maize are the main staple food crops to ensure the food security in China with diversified climate condition, cropping system and environmental and socio-economic factors across provinces. Spatial variation of technical efficiency in farmers’ field is helpful to understand the potential to improve farmers’ yield given the inputs level and reduce the yield gap. The study is based on a large-scale farm household survey which covered 1 218 rice farmers, 3 566 wheat farmers and 2 111 maize farmers in the main producing areas. The results indicate that rice farmers are with very high technical efficiency level, nearly 0.9 on average, with little room to improve the efficiency of agricultural inputs. Similar results have been found in wheat and maize farmers’ fields, although the technical efficiency levels are lower than that of rice farmers while still at a high level with obvious variation across regions. Farmers with higher yield level also achieve better technical efficiency in most locations. Both local environmental and socio-economic factors significantly affect farmers’ technical efficiency. In the context of urbanization and economic development, improved and new agricultural technologies need to be prioritized and facilitated to improve cereal yield at farm level.

High-performance liquid chromatography (HPLC) was employed to determine drug release rates based on emamectin benzoate concentrations in the medium.  Release kinetics equations were used to fit the drug release behavior.  The effects of particle size and release medium pH on the release rate were also investigated.  The indoor toxicity of emamectin benzoate-loaded polylactic acid microspheres on the diamondback moth larva (Plutella xylostella) was studied to explore drug sustained-release performance.  In acidic and neutral media, the drug release behavior of the microspheres was in accord with the first-order kinetics equation.  Increasing the spray dosage of emamectin benzoate-loaded polylactic acid microspheres initially resulted in an equivalent insecticidal efficacy with the conventional emamectin benzoate microemulsion.  However, the drug persistence period was four-fold longer than that observed using the conventional formulation.  The developed emamectin benzoate-loaded polylactic acid microspheres showed dramatic sustained-release performance.  A treatment threshold of greater than 35 mg mL^–1 was established for an efficient accumulated release concentration of emamectin benzoate-loaded microspheres.

Blast resistance and grain quality are major problems in hybrid rice production in China.  In this study, two resistance (R) genes, Pi46 and Pita, along with the gene Wx^b, which mainly affects rice endosperm amylose content (AC), were introgressed into an elite indica restoring line, R8166, which has little blast resistance and poor grain quality through marker-assisted selection (MAS).  Eight improved lines were found to have recurrent genome recovery ratios ranging from 88.68 to 96.23%.  Two improved lines, R163 and R167, were selected for subsequent studies.  R167, which has the highest recovery ratio (96.23%), showed no significant differences in multiple agronomic traits.  In contrast, R163 with the lowest recovery ratio (88.68%) exhibited significant differences in heading date and yield per plant compared with the recurrent parent.  At two developmental stages, R163 and R167 had greatly enhanced resistance to blast over the recurrent parent.  Similar trends were also observed for agronomic traits and blast resistance in R163- and R167-derived hybrids when compared with the counterparts from R8166.  In addition, R163, R167, and their derived hybrids significantly improved the grain quality traits, including amylose content (AC), gel consistency (GC), chalky grain rate (CGR), and degree of endosperm chalkiness (DEC).  It confirmed the success of efficiently developing elite restoring lines using MAS in this study.

    Homeobox 1 in Malus×domestica (MdHB-1) is a transcription factor that belongs to homeodomain-leucine zipper I (HD-Zip I) protein subfamily. According to previous reports, MdHB-1 could regulate ethylene synthesis by binding with the MdACO1 promoter, but other functions of MdHB-1 are still unknown. To reveal more clues concerning the characters of the MdHB-1 gene promoter and the functions of MdHB-1, the promoter region of MdHB-1 was cloned from the Royal Gala apple genome and recombined with the β-glucuronidase (GUS) gene in this study. This research was conducted in Nicotiana tabacum and supported by Agrobacterium-mediated transient transformation and bioinformatics analysis. Deletion analysis of the MdHB-1 promoter showed that the GUS gene could be activated by serially deleted promoters, and the activity promoted by 680 nucleotides (nt) was the lowest. The region, which is 266 nt upstream of the initiation code (ATG), was effective for GUS expression. Meanwhile, the activity of the MdHB-1 promoter (-1 057 nt), which was stronger than MdHB-1 promoter (-1 057 to -266 nt) and lack the 5´-untranslated region (5´-UTR), showed that 5´-UTR may have a positive effect on gene transcription. After the sequence analysis, the cis-acting elements that respond to hormones and environmental stresses were identified in the promoter region. The MdHB-1 promoter (1 057 nt) activity in Nicotiana tabacum was positively induced by ethrel and darkness, and it was suppressed by gibberellic acid (GA), whereas abscisic acid (ABA), salicylic acid (SA), wounding, and Pseudomonas syringae pv. tomato (DC3000) treatments revealed a slight auxo-action. These results reveal that the MdHB-1 promoter receive internal or external signals, and MdHB-1 may refer to many biological activities in apple, such as its stress response, development, and ripening.

    With increased cultivation of transgenic Bacillus thuringiensis (Bt) cotton in the saline alkaline soil of China, assessments of transgenic crop biosafety have focused on the effects of soil salinity on rhizosphere microbes and Bt protein residues. In 2013 and 2014, investigations were conducted on the rhizosphere microbial biomass, soil enzyme activities and Bt protein contents of the soil under transgenic Bt cotton (variety GK19) and its parental non-transgenic cotton (Simian 3) cultivated at various salinity levels (1.15, 6.00 and 11.46 dS m⁻¹). Under soil salinity stress, trace amounts of Bt proteins were observed in the Bt cotton GK19 rhizosphere soil, although the protein content increased with cotton growth and increased soil salinity levels. The populations of slight halophilic bacteria, phosphate solubilizing bacteria, ammonifying bacteria, nitrifying bacteria and denitrifying bacteria decreased with increased soil salinity in the Bt and non-Bt cotton rhizosphere soil, and the microbial biomass carbon, microbial respiration and soil catalase, urease and alkaline phosphatase activity also decreased. Correlation analyses showed that the increased Bt protein content in the Bt cotton rhizosphere soil may have been caused by the slower decomposition of soil microorganisms, which suggests that salinity was the main factor influencing the relevant activities of the soil microorganisms and indicates that Bt proteins had no clear adverse effects on the soil microorganisms. The results of this study may provide a theoretical basis for risk assessments of genetically modified cotton in saline alkaline soil.

Nutrient deficiency stresses often occur simultaneously in soil.  Thus, it’s necessary to investigate the mechanisms underlying plant responses to multiple stresses through identification of some key stress-responsive genes.  Quantitative real-time PCR (qRT-PCR) is essential for detecting the expression of the interested genes, of which the selection of suitable reference genes is a crucial step before qRT-PCR.  To date, reliable reference genes to normalize qRT-PCR data under different nutrient deficiencies have not been reported in plants.  In this study, expression of ten candidate reference genes was detected in leaves and roots of rapeseed (Brassica napus L.) after implementing different nutrient deficiencies for 14 days.  These candidate genes, included two traditionally used reference genes and eight genes selected from an RNA-Seq dataset.  Two software packages (GeNorm, NormFinder) were employed to evaluate candidate gene stability.  Results showed that VHA-E1 was the highest-ranked gene in leaves of nutrient-deficient rapeseed, while VHA-G1 and UBC21 were most stable in nutrient-deficient roots.  When rapeseed leaves and roots were combined, UBC21, HTB1, VHA-G1 and ACT7 were most stable among all samples.  To evaluate the stabilities of the highest-ranked genes, the relative expression of two target genes, BnTrx1;1 and BnPht1;3 were further determined.  The results showed that the relative expression of BnTrx1;1 depended on reference gene selection, suggesting that it’s necessary to evaluate the stability of reference gene prior to qRT-PCR.  This study provides suitable reference genes for gene expression analysis of rapeseed responses to different nutrient deficiencies, which is essential for elucidation of mechanisms underlying rapeseed responses to multiple nutrient deficiency stresses.

    Studies were conducted to identify candidate soil microbes responsible for observed differences in strawberry vigour at a small spatial scale, which was not associated with visual disease symptoms. Samples were obtained from the soils close to the rhizosphere of ‘big’ and ‘small’ plants from small plots which exhibited large local heterogeneity in plant vigour. A metabarcoding approach was used to profile bacterial and fungal compositions, using two primer pairs for 16S ribosomal RNA genes (16S rDNA) and one for the fungal internal transcribed spacer (ITS) region. Of the two 16S rDNA primer sets, the 341F/805R resulted in sequences of better quality. A total 28 operational taxonomic units (OTUs) had differential relative abundance between samples from ‘big’ and ‘small’ plants. However, plausible biological explanation was only possible for three fungal OTUs. Two were possible phytopathogens: Verticillium spp. and Alternaria alternata although the latter has never been considered as a main pathogen of strawberry in the UK. For samples from ‘small’ plants, the abundance of these OTUs was much greater than from ‘big’ plants. The opposite was true for a mycorrhizal OTU. These results suggest that soil microbes related to crop production can be identified using metabarcoding technique. Further research is needed to assess whether A. alternata and Verticillium spp. could affect strawberry growth in the field.

    The structural characteristics of protein body accumulation in different endosperm regions of hard wheat cultivar (XM33) and soft wheat cultivar (NM13) under drought stress were investigated. Drought stress treatment was implemented from plant regreening to the caryopsis mature stage. Microscope images of endosperm cells were obtained using resin semi-thin slice technology to observe the distribution and relative area of protein body (PB). Compared with NM13, relative PB area of XM33 was significantly higher in sub-aleurone endosperm region. The amount of accumulation, including the size and relative area of PB, in two wheat cultivars was higher in sub-aleurone region than that in central region at 18 days post anthesis (DPA). Drought stress significantly enhanced the sizes and relative areas of PBs in the dorsal and abdominal endosperms in two wheat cultivars. Particularly for dorsal endosperm, drought stress enhanced the relative PB area at 18 DPA and NM13 (5.0% vs. 6.73%) showed less enhancement than XM33 (5.49% vs. 8.96%). However, NM13 (9.58% vs. 12.02%) showed greater enhancement than XM33 (10.25% vs. 11.7%) at 28 DPA. The protein content in the dorsal and abdominal endosperms of the two wheat cultivars decreased at 12 DPA and then increased until 38 DPA. Drought stress significantly increased the protein contents in the two main regions. From 12 to 38 DPA, the amount of PB accumulation and the protein content were higher in XM33 than those in NM13. The results revealed that PB distribution varied in different endosperm tissues and that the amount of PB accumulation was remarkably augmented by drought stress.

    Utilization of R (resistance) genes to develop resistant cultivars is an effective strategy to combat against rice blast disease. In this study, R genes Pi46 and Pita in a resistant accession H4 were introgressed into an elite restorer line Hang-Hui-179 (HH179) using the marker-assisted backcross breeding (MABB) procedure. As a result, three improved lines (e.g., R1791 carrying Pi46 alone, R1792 carrying Pita alone and R1793 carrying both Pi46 and Pita) were developed. The three improved lines had significant genetic similarities with the recurrent parent HH179. Thus, they and HH179 could be recognized as near isogenic lines (NILs). The resistance spectrum of the three improved lines, which was tested at seedling stage, reached 91.1, 64.7 and 97.1%, respectively. This was markedly broader than that of HH179 (23.5%). Interestingly, R1793 showed resistance to panicle blast but neither R1791 nor R1792 exhibited resistance at two natural blast nurseries. The results implied that the stacking of Pi46 and Pita resulted in enhanced resistance, which was unachievable by either R gene alone. Further comparison indicated that the three improved lines were similar to HH179 in multiple agronomic traits; including plant height, tillers per plant, panicle length, spikelet fertility, and 1 000-grain weight. Thus, the three improved lines with different R genes can be used as new sources of resistance for developing variety. There is a complementary effect between the two R genes Pi46 and Pita.