GsMAPK4, a positive regulator of soybean tolerance to salinity stress

Salt stress is one of the major factors affecting plant growth and yield in soybean under saline soil condition. Despite many studies on salinity tolerance of soybean during the past few decades, the detailed signaling pathways and the signaling molecules for salinity tolerance regulation have not been clarified. In this study, a proteomic technology based on two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) were used to identify proteins responsible for salinity tolerance in soybean plant. Real-time quantitative PCR (qRT-PCR) and Western blotting (WB) were used to verify the results of 2-DE/MS. Based on the results of 2-DE and MS, we selected glucosyltransferase (GsGT4), 4-coumarate, coenzyme A ligase (Gs4CL1), mitogen-activated protein kinase 4 (GsMAPK4), dehydration responsive element binding protein (GsDREB1), and soybean cold-regulated gene (GsSRC1) in the salinity tolerant soybean variety, and GsMAPK4 for subsequent research. We transformed soybean plants with mitogen-activated-protein kinase 4 (GsMAPK4) and screened the resulting transgenics soybean plants using PCR and WB, which confirmed the expression of GsMAPK4 in transgenic soybean. GsMAPK4-overexpressed transgenic plants showed significantly increased tolerance to salt stress, suggesting that GsMAPK4 played a pivotal role in salinity tolerance. Our research will provide new insights for better understanding the salinity tolerance regulation at molecular level.     

Evaluating effective Trichoderma isolates for biocontrol of Rhizoctonia solani causing root rot of Vigna unguiculata

The highly diverse genus Trichoderma has provided many formulations that are alternatives to the chemical pesticides in agriculture. The present study was undertaken to investigate the biocontrol potential of eight Trichoderma species, T. atrobrunneum, T. guizhouense, T. paratroviride, T. pyramidale, T. rufobrunneum, T. simmonsii, T. thermophilum and T. viridulum, against the phytopathogenic fungus Rhizoctonia solani. Trichoderma isolates were first evaluated in vitro by dual culture tests for their antagonism, mycoparasitic ability and antifungal activity against R. solani. Their growth promoting potential was further assessed in relation to phosphate solubilization, indole acetic acid and siderophore production. Five of the isolates were selected and evaluated for their abilities to prompt plant growth and to control R. solani infecting Vigna unguiculata(cowpea) seedlings in vivo. Two most effective isolates, T. guizhouense 9185 and T. simmonsii 8702, significantly(P0.05) reduced the disease severity incidences(36.6 and 45.0%, respectively) and promoted plant growth, which have good prospects for application.     

Estimates on nitrogen uptake in the subsequent wheat by above-ground and root residue and rhizodeposition of using peanut labeled with 15N isotope on the North China Plain

Leguminous crops play a vital role in enhancing crop yield and improving soil fertility. Therefore, it can be used as an organic N source for improving soil fertility. The purpose of this study was to (i) quantify the amounts of N derived from rhizodeposition, root and above-ground biomass of peanut residue in comparison with wheat and (ii) estimate the effect of the residual N on the wheat-growing season in the subsequent year. The plants of peanut and wheat were stem fed with ¹⁵N urea using the cotton-wick method at the Wuqiao Station of China Agricultural University in 2014. The experiment consisted of four residue-returning strategies in a randomized complete-block design: (i) no return of crop residue (CR0); (ii) return of above-ground biomass of peanut crop (CR1); (iii) return of peanut root biomass (CR2); and (iv) return of all residue of the whole peanut plant (CR3). The 31.5 and 21% of the labeled ¹⁵N isotope were accumulated in the above-ground tissues (leaves and stems) of peanuts and wheat, respectively. N rhizodeposition of peanuts and wheat accounted for 14.91 and 3.61% of the BG¹⁵N, respectively. The ¹⁵N from the below-ground ¹⁵N-labeled of peanuts were supplied 11.3, 5.9, 13.5, and 6.1% of in the CR0, CR1, CR2, and CR3 treatments, respectively. Peanut straw contributes a significant proportion of N to the soil through the decomposition of plant residues and N rhizodeposition. With the current production level on the NCP, it is estimated that peanut straw can potentially replace 104 500 tons of synthetic N fertilizer per year. The inclusion of peanut in rotation with cereal can significantly reduce the use of N fertilizer and enhance the system sustainability.     

Molecular identification and enzymatic properties of laccase2 from the diamondback moth Plutella xylostella (Lepidoptera: Plutellidae)

Laccase(EC 1.10.3.2)is known to oxidize various aromatic and nonaromatic compounds via a radical-catalyzed reaction,which generally includes two types of laccase,Lac1 and Lac2.Lac1 oxidizes toxic compounds in the diet,and Lac2 is known to play an important role in melanizing the insect exoskeleton.In this study,we cloned and sequenced the cDNA of the diamondback moth,Plutella xylostella Lac2(PxLac2),from the third instar larvae using polymerase chain reaction(PCR)and rapid amplification of cDNA ends techniques.The results showed that the full-length PxLac2 cDNA was 1 944 bp long and had an open reading frame of 1 794 bp.PxLac2 encoded a protein with 597 amino acids and had a molecular weight of 66.09 kDa.Moreover,we determined the expression levels of PxLac2 in different stages by quantitative PCR(qPCR).The results indicated that PxLac2 was expressed differently in different stages.We observed the highest expression level in pupae and the lowest expression level in fourth instar larvae.We also investigated the enzymatic properties of laccase,which had optimal activity at pH 3.0 and at 35°C.Under these optimal conditions,laccase had a Michaelis constant(K_m)of 0.97 mmol L~(-1),maximal reaction speed(V_m)of 56.82 U mL~(-1),and activation energy(E_a)of 17.36 kJ mol~(-1) to oxidize2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid ammonium salt).Type II copper enhanced laccase activity below 0.8mmol L~(-1) and reduced enzyme activity above 0.8 mmol L~(-1) with an IC_(50) concentration of 1.26 mmol L~(-1).This study provides insights into the biological function of laccase.     

Phenotypic characterization and genetic mapping of the dwarf mutant m34 in maize

Plant height is one of the most important agronomic traits associated with yield in maize.In this study,a gibberellins(GA)-insensitive dwarf mutant,m34,was screened from inbred line Ye478 by treatment with the chemical mutagen ethylmethanesulfonate(EMS).Compared to Ye478,m34 showed a dwarf phenotype with shorter internodes,and smaller leaf length and width,but with similar leaf number.Furthermore,m34 exhibited smaller guard cells in internodes than Ye478,suggesting that smaller cells might contribute to its dwarf phenotype.Genetic analysis indicated that the m34 dwarf phenotype was controlled by a recessive nuclear gene.An F2 population derived from a cross between m34 and B73 was used for mutational gene cloning and this gene was mapped to a chromosome region between umc2189 and umc1553 in chromosome 1 bin1.10,which harbored a previously identified dwarf gene Zm VP8.Sequencing analysis showed a nucleotide substitution(G1606 to A1606)in the sixth exon of ZmVP8,which resulted in an amino acid change(E531 to K531)from Ye478 to m34.This amino acid change resulted in anα-helix changing to aβ-sheet in the secondary protein structure and the‘SPEC’domain changed to a‘BOT1NT’domain in the tertiary protein structure.Taken together,these results suggested that m34 is a novel allelic mutant originally derived from Ye478 that is useful for further ZmVP8 functional analysis in maize.     

The synergistic advantage of combining chloropicrin or dazomet with fosthiazate nematicide to control root-knot nematode in cucumber production

The highly-damaging root-knot nematode(Meloidogyne spp., RKN) cannot be reliably controlled using only a nematicide such as fosthiazate because of increasing pest resistance. In laboratory and greenhouse trials, we showed that chloropicrin(CP) or dazomet(DZ) synergized the efficacy of fosthiazate against RKN. The combination significantly extended the degradation half-life of fosthiazate by an average of about 1.25 times. CP or DZ with fosthiazate reduced the time for fosthiazate to penetrate the RKN cuticle compared to fosthiazate alone. CP or DZ combined with low or medium rate of fosthiazate increased the total cucumber yield, compared to the use of each product alone. A low-dose fosthiazate with DZ improved total yield more than a low dose fosthiazate with CP. Extending the half-life of fosthiazate and reducing the time for fosthiazate or fumigant to penetrate the RKN cuticle were the two features that gave the fumigant-fosthiazate combination its synergistic advantage over these products used singularly. This synergy provides the opportunity for farmers to use a low dose of fosthiazate which lowers the risk of RKN resistance. Farmers could combine DZ at 30 g m~(-2) with fosthiazate at a low rate of 0.375 g m~(-2) to control RKN and adequately control two major soil-borne diseases in cucumber greenhouses.     

Genetic dissection of hexanol content in soybean seed through genome-wide association analysis

Hexanol is a major compound contributing to the off-flavors(the bean-like odor) of soybean derived soymilk. The most effective way to reduce the off-flavors of soymilk is the screening and utilization of soybean cultivars with improved hexanol content. However, no genome-wide genetic analysis for this particular trait has been conducted to date. The objective of the present study was to dissect the genetic basis of hexanol content in soybean seed through genome-wide association analysis(GWAS). A total of 105 soybean accessions were analyzed for hexanol content in a three-year experiments and genotyped by sequencing using the specific locus amplified fragment sequencing(SLAF-seq) approach. A total of 25 724 single nucleotide polymorphisms(SNPs) were obtained with minor allele frequencies(MAF)5%. GWAS showed that 25 quantitative trait nucleotides(QTNs) were significantly associated with the hexanol concentration in soybean seed. These identified QTNs distributed on different genomic regions of the 15 chromosomes. A total of 91 genes were predicted as candidate genes underlying the seed hexanol level and six candidates were predicted possibly underlying the seed hexanol by gene based association. In this study, GWAS has been proven to be an effective way to dissect the genetic basis of the hexanol concentration in multiple genetic backgrounds. The identified beneficial alleles and candidate genes might be valuable for the improvement of marker-assisted breeding efficiency for low hexanol level and help to explore possible molecular mechanisms underlying hexanol content in soybean seed.     

Molecular,serological and biological characterization of a novel Apple stem pitting virus strain from a local pear variety grown in China

Apple stem pitting virus (ASPV) is an important causal agent of pear diseases. Nowadays, the infection status and molecular characteristics of the virus in old pear trees have never been investigated. In this study, we provide the first complete genome sequence of an ASPV isolate LYC from an over 300-year-old tree of a local Pyrus bretschneideri cultivar ‘Chili’ specifically grown at Laiyang area in China. ASPV-LYC possesses a chimeric genome consisting of 9 273 nucleotides excluding a poly(A) tail at its 3’ end and harboring a recombination region in its open reading frame (ORF1) with Aurora-1 and KL9 identified as the major and minor parents. Western blot analysis with antisera against recombinant coat proteins (CPs) of three ASPV isolates from pear indicates that ASPV-LYC is serologically related to these ASPV isolates, but with differential activities. Further biological tests on indicator plants of Pyronia veitchii show that ASPV-LYC can induce serious leaf and stem symptoms as other ASPV isolates. The results provide an important information for understanding molecular evolution of ASPV and suggest a need to prevent dissemination of the isolate among pear trees.     

Characterization of TaCOMT genes associated with stem lignin content in common wheat and development of a gene-specific marker

Stem lignin content(SLC) in common wheat(Triticum aestivum L.) contributes to lodging resistance. Caffeic acid 3-O-methyltransferase(COMT) is a key enzyme involved in lignin biosynthesis. Characterization of TaCOMT genes and development of gene-specific markers could enable marker-assisted selection in wheat breeding. In the present study, the full-length genomic DNA(gDNA) sequences of TaCOMT genes located on chromosomes 3 A, 3 B, and 3 D were cloned by homologous cloning. Two allelic variants, TaCOMT-3 Ba and TaCOMT-3 Bb, were identified and differed by a 222-bp insertion/deletion(InDel) in the 3′-untranslated region(3′-UTR). A co-dominant gene-specific marker based on this InDel was developed and designated as Ta COMT-3 BM. A total of 157 wheat cultivars and advanced lines grown in four environments were used to validate the associations between allelic patterns and SLC. The SLC of cultivars with TaCOMT-3 Ba was significantly(P0.01) higher than that of those with TaCOMT-3 Bb, and the marker TaCOMT-3 BM could be effectively used in wheat breeding.     

Reduction in cadmium accumulation in japonica rice grains by CRISPR/Cas9-mediated editing of OsNRAMP5

Cadmium (Cd) intake is harmful to human health and Cd contamination in rice grains represents a severe threat to those consuming rice as a staple food. Knockout of Cd transporters is a promising strategy to reduce Cd accumulation in rice grains. OsNRAMP5 is the major transporter for Cd and manganese (Mn) uptake in rice. Nevertheless, it is uncertain whether knockout of OsNRAMP5 is applicable to produce low Cd rice without affecting plant growth and grain yield. In this study, we adopted CRISPR/Cas9-based gene editing technology to knock out OsNRAMP5 in two japonica varieties. We generated three independent transgene-free osnramp5 mutants and investigated the effect of osnramp5 mutations on Cd accumulation and plant growth. Hydroponic experiments showed that plant growth and chlorophyll content were significantly reduced in osnramp5 mutants at low Mn conditions, and this defective growth in the mutants could be fully rescued by supply of high levels of Mn. Cd and Mn accumulation in both roots and shoots was markedly reduced in the mutants compared to that in wild-type plants. In paddy field experiments, although Cd in flag leaves and grains was greatly reduced in osnramp5 mutants, some agronomic traits including plant height, seed setting rate, and grain number per panicle were affected in the mutants, which ultimately caused a mild reduction in grain yield. The reduced plant growth in the mutants can be attributed to a marked decrease in Mn accumulation. Our results reveal that the manipulation of OsNRAMP5 should be treated with caution: When assessing the applicability of osnramp5 mutants, soil pH and soil water content in paddy fields need to be taken into consideration, since they might affect the levels of available Mn in the soil and consequently determine the effect of the mutation on grain yield.     

Molecular cloning and functional characterization of apple U-box E3 ubiquitin ligase gene MdPUB29 reveals its involvement in salt tolerance

An E3 ubiquitin ligase gene(Genbank accession no.: MD01 G1010900) was cloned from the Royal Gala apple genome(Malus×domestica Borkh.). Sequence analysis showed that the length of the MdPUB29 gene was 1 275 bp, encoding 424 amino acids. Phylogenetic tree analysis indicated that the apple E3 ubiquitin ligase exhibited the greatest sequence similarity to Pyrus×bretschneideri. The predicted protein structural domain of MdPUB29 showed that it contained a U-box domain. qRT-PCR analysis showed that Md PUB29 was expressed widely in different tissues of the Royal Gala apple species, and was highly expressed in the root, while the expression of MdPUB29 was significantly inhibited by exogenous NaCl. Immunoblotting assays revealed that MdPUB29 protein abundance in tissue cultures of the Royal Gala apple accumulated under NaC l stress conditions. Three-dimensional protein structure prediction indicated that MdPUB29 was highly homologous with AtPUB29. The growing potential of MdPUB29-expressing apple calli and Arabidopsis were much stronger than that of the control under salt stress conditions, suggesting that MdPUB29 may positively regulate salt tolerance.     

The effects of soil moisture and salinity as functions of groundwater depth on wheat growth and yield in coastal saline soils

In the coastal saline soils, moisture and salinity are the functions of groundwater depth affecting crop growth and yield. Accordingly, the objectives of this study were to: 1) investigate the combined effects of moisture and salinity stresses on wheat growth as affected by groundwater depth, and 2) find the optimal groundwater depth for wheat growth in coastal saline soils. The groundwater depths (0.7, 1.1, 1.5, 1.9, 2.3, and 2.7 m during 2013–2014 (Y1) and 0.6, 1.0, 1.4, 1.8, 2.2, and 2.6 m during 2014–2015 (Y2)) of the field experiment were maintained by soil columns. There was a positive correlation between soil moisture and salinity. Water logging with high salinity (groundwater depth at 0.7 m in Y1 and 0.6 m in Y2) showed a greater decline towards wheat growth than that of slight drought with medium (2.3 m in Y1) or low salinity (2.7 m in Y1, 2.2 and 2.6 m in Y2). The booting stage was the most sensitive stage of wheat crop under moisture and salinity stresses. Data showed the most optimal rate of photosynthesis, grain yield, and flour quality were obtained under the groundwater depth (ditch depth) of 1.9 m (standard soil moisture with medium salinity) and 2.3 m (slight drought with medium salinity) in Y1 and 1.8 m (standard soil moisture with medium salinity) and 2.2 m (slight drought with low salinity) in Y2. The corresponding optimal soil relative moisture content and conductivity with the 1:5 distilled water/soil dilution, in the depth of 0–20 cm and 20–40 cm in coastal saline soils, were equal to 58.67–63.07% and 65.51–72.66% in Y1, 63.09–66.70% and 69.75–74.72% in Y2; 0.86–1.01 dS m^–1 and 0.63–0.77 dS m^–1 in Y1, 0.57–0.93 dS m^–1 and 0.40–0.63 dS m^–1 in Y2, respectively.     

Species-specific COI primers for rapid identification of a globally significant invasive pest,the cassava mealybug Phenacoccus manihoti Matile-Ferrero

The globally invasive cassava mealybug Phenacoccus manihoti Matile-Ferrero is a pernicious pest of cassava,and its recent introduction into Asia has raised considerable alarm.To slow or prevent further invasion,an accurate,simple,and developmental-stage-independent detection method for P.manihoti is required.In the present study,a PCR method based on a species-specific mitochondrial DNA cytochrome oxidase I(SS-COI)marker was developed for rapid identification of P.manihoti.One pair of SS-COI primers(PMSSZW-1F and PMSSZW-1R)was designed based on sequence variations in the COI gene among P.manihoti and related mealybug species.Specificity of the primer pair was validated on 21 closely related species.Sensitivity tests were performed on four immature developmental stages and female adults.Efficacy tests demonstrated that at the relatively low concentration of(135.2±14.7)pgresuspended DNA,the specific fragment was detected in all replicates.Furthermore,the SS-COI primer pair was assayed on three populations of P.manihoti from major exporting countries of cassava.The PCR assay was proved to be a rapid,simple,and reliable molecular measure for the identification of P.manihoti.This tool will be useful for quarantine,monitoring,and management of this invasive pest.     

Effects of the severity and timing of basal leaf removal on the amino acids profiles of Sauvignon Blanc grapes and wines

The effects of the severity and timing of leaf removal(LR) on the amino acids of Sauvignon Blanc grapes and wines were studied during the 2017 growing season. High-performance liquid chromatography(HPLC) was used to analyze the amino acids profiles of grape berries and wines. The basal leaves were removed at three time points(40, 56 and 72 days after flowering, named LR40, LR56 and LR72, respectively) at two severity levels(one at which the first, third, and fifth basal leaves of each shoot were removed(50% level); and another at which the first six basal leaves were removed(100% level)). The results showed that leaf removal had little impact on total soluble solids(°Brix), titratable acidity, pH or berry weight. The LR72-50% treated grapes had higher berry weight, titratable acidity and °Brix than those of the other treatments. The highest concentrations of total amino acids and of total amino acids except proline were detected in LR72-50% treated grapes(2 952.58 and 2 764.36 mg L~(-1), respectively); the lowest were detected in LR72-100% treated grapes(2 172.82 and 2 038.71 mg L~(-1), respectively). LR72-50% treatment significantly promoted the synthesis of aspartic acid, serine, arginine, alanine, aminobutyric acid and proline at both severity levels for grapes, the concentrations of all of these amino acids were increased relative to the control concentrations. The LR72-50%, LR40-100% and LR72-100% treated wines had higher total amino acids concentrations and higher concentrations of some individual amino acids, such as arginine, alanine and serine, than did the control wines. Of all the amino acids studied, glycine, tyrosine, cysteine, methionine and lysine were not significantly influenced by the timing or severity basal defoliation in grapes and wines. The present study reveals the effects of the timing and severity of leaf removal on the amino acids profiles of grapes and wines.     

Mitochondrial DNA diversity and origin of indigenous pigs in South China and their contribution to western modern pig breeds

Indigenous pigs in South China are valuable genetic resources with many specific and unique characters, which have played an important role in the establishment of some western modern pig breeds. However, the origin and genetic diversity of indigenous pigs in South China have not been fully understood. In the present study, we sequenced 534 novel mitochondrial DNA(mt DNA) D-loop and assembled 54 complete mitogenome sequences for all 17 indigenous pig breeds from Fujian, Guangdong, Guangxi and Hainan in South China. These data were analyzed together with previously published homologous sequences relevant to this study. We found that all 13 coding genes of the mitogenomes were under purifying selection, but ND1 had the most variable sites and CYTB contained the most non-synonymous SNPs. Phylogenetic analysis showed that all indigenous pigs in South China were clustered into the D haplogroup with D1 a1, D1 b, D1 c and D1 e sub-haplogroups found to be dominant. Haplotype and nucleotide diversities of D-loop sequences ranged from 0.427 to 0.899 and from 0.00342 to 0.00695, respectively, among which all pigs in Guangdong had the lowest diversity. The estimates of pairwise FST, gene flow(Nm) and genetic distance(Da) indicated that most of these indigenous pig breeds differentiated from each other significantly(P0.05). Among the western modern breeds, Berkshire and Yorkshire had significant Asian matrilineal footprints from indigenous pigs in South China, especially the Spotted pigs distributed in Guangdong and Guangxi. The neutrality test(Fu's F_S) indicated that indigenous pigs from Fujian and Guangxi had gone through recent population expansion events(P0.05). It is concluded that indigenous pigs in South China were most likely derived from the Mekong region and the middle and downstream regions of Yangtze River through Guangxi and Fujian. Our findings provide a complete and in-depth insight on the origin and distribution pattern of maternal genetic diversity of indigenous pigs in South China.     

First detection and complete genome of Soybean chlorotic mottle virus naturally infecting soybean in China by deep sequencing

Soybean chlorotic mottle virus (SbCMV) was first detected from soybean plants in Jiangxi Province of China by high throughput sequencing and was confirmed by PCR. The complete nucleotide sequence of NC113 was determined to be 8 210 nucleotides, and shared the highest similarity (91.7%) with sequences of SbCMV that was only reported in Japan. It encodes nine putative open reading frames (ORFs Ia, Ib and II–VIII), and contains a large intergenic region located at nucleotide 5 976–6 512 between ORFs VI and VII. Sequence analysis and phylogenetic tree indicated that NC113 is an isolate of SbCMV, and is more related to the soymoviruses Blueberry red ringspot virus (BRRSV), Peanut chlorotic streak virus (PCSV) and Cestrum yellow leaf curling virus (CmYLCV) than to other representative members in the Caulimoviridae family. Field survey of 472 legume plants from Jiangxi and Zhejiang provinces showed SbCMV was only detected from soybean in Nanchang City with a low incidence rate. This is the first report of Soybean chlorotic mottle virus identified in China.     

Combined effect of shading time and nitrogen level on grain filling and grain quality in japonica super rice

There is limited information about the combined effect of shading time and nitrogen(N) on grain filling and quality of rice. Therefore, two japonica super rice cultivars, Nanjing 44 and Ningjing 3, were used to study the effect of shading time and N level on the characteristics of rice panicle and grain filling as well as the corresponding yield and quality. At a low N level(150 kg N ha~(–1), 150 N), grain yield decreased(by 21.07–26.07%) under the treatment of 20 days of shading before heading(BH) compared with the no shading(NS) treatment. These decreases occurred because of shortened panicle length, decreased number of primary and secondary branches, as well as the grain number and weight per panicle. At 150 N, in the treatment of 20 days of shading after heading(AH), grain yield also decreased(by 9.46–10.60%) due to the lower grain weight per panicle. The interaction of shading and N level had a significant effect on the number of primary and secondary branches. A high level of N(300 kg N ha~(–1), 300 N) could offset the negative effect of shading on the number of secondary branches and grain weight per panicle, and consequently increased the grain yield in both shading treatments. In superior grains, compared with 150 N NS, the time to reach 99% of the grain weight(T_(99)) was shortened by 1.6 to 1.7 days, and the grain weight was decreased by 4.18–5.91% in 150 N BH. In 150 N AH, the grain weight was 13.39–13.92% lower than that in 150 N NS due to the slow mean and the maximum grain-filling rate(GR_(mean )and GR_(max)). In inferior grains, grain weight and GR_(mean) had a tendency of 150 N NS150 N BH150 N AH. Under shaded conditions, 300 N decreased the grain weight due to lower GR_(mean) both in superior and inferior grains. Compared with 150 N NS, the milling and appearance qualities as well as eating and cooking quality were all decreased in 150 N BH and 150 N AH. Shading with the high level of 300 N improved the milling quality and decreased the number of chalky rice kernels, but the eating and cooking quality was reduced with increased chalky area and overall chalkiness. Therefore, in the case of short term shading, appropriate N fertilizer could be used to improve the yield and milling quality of rice, but limited application of N fertilizer is recommended to achieve good eating and cooking quality of rice.