Targeted mutations of BnPAP2 lead to a yellow seed coat in Brassica napus L.

The yellow seed trait is preferred by breeders for its potential to improve the seed quality and commercial value of Brassica napus.In the present study,we produced yellow seed mutants using a CRISPR/Cas9 system when the two BnPAP2 homologs were knocked out.Histochemical staining of the seed coat demonstrated that proanthocyanidin accumulation was significantly reduced in the pap2 double mutants and decreased specifically in the endothelial and palisade layer cells of the seed coat.Transcriptomic and metabolite profiling analysis suggested that disruption of the BnPAP2 genes could reduce the expression of structural and regulated genes in the phenylpropanoid and flavonoid biosynthetic pathways.The broad suppression of these genes might hinder proanthocyanidin accumulation during seed development,and thereby causing the yellow seed trait in B.napus.These results indicate that BnPAP2 might play a vital role in the regulatory network controlling proanthocyanidin accumulation.

Fine-mapping and transcriptome analysis of the photosensitive leaf-yellowing gene CaLY1 in pepper(Capsicum annuum L.)

Leaf color is directly related to altered photosynthesis.Hence,leaf yellowing mutants can be widely used for the researching plant physiology and functional genomes,for cultivating new varieties of popular horticultural plants,and for identifying hybrid purity(as markers).Here,we constructed a^(60)Co-γF_(2)population from the leaf-yellowing mutant R24 via radiation mutation with the inbred line WT21 of pepper.Genetic analysis showed that the leaf-yellowing of the mutant was controlled by a single recessive gene.By applying the Bulk Segregation Analysis and Kompetitive Allele Specific PCR markers,the leaf-yellowing gene CaLY1(Capsicum annuum Leaf yellow 1)was mapped on chromosome 9,SNP5791587-SNP6011215,with a size of 214.5 kb.One non-synonymous mutated gene Capana09g000166 was found in the interval.The gene encoded a Psb X,which is the core complex of PSⅡ.Transcriptome analysis further showed that 2301 differentially expressed genes were identified under shading treatment for 24 h in R24.The Gene Ontology enrichment pathways were related to photosynthesis light harvesting,cell wall,activity of quercetin 3-O-glucosyltransferase and flavonoid metabolic process,which likely regulate the response of pepper leaves to different light levels.Functional enrichment analysis indicated that the most abundant pathways were photosynthesis antenna proteins and metabolic.

Grain Shape Genes:Shaping the Future of Rice Breeding

The main goals of rice breeding nowadays include increasing yield,improving grain quality,and promoting complete mechanized production to save labor costs.Rice grain shape,specified by three dimensions,including grain length,width and thickness,has a more precise meaning than grain size,contributing to grain appearance quality as well as grain weight and thus yield.Furthermore,the divergence of grain shape characters could be utilized in mechanical seed sorting in hybrid rice breeding systems,which has been succeeded in utilizing heterosis to achieve substantial increase in rice yield in the past decades.Several signaling pathways that regulate rice grain shape have been elucidated,including G protein signaling,ubiquitination-related pathway,mitogen-activated protein kinase signaling,phytohormone biosynthesis and signaling,micro RNA process,and some other transcriptional regulatory pathways and regulators.This review summarized the recent progress on molecular mechanisms underlying rice grain shape determination and the potential of major genes in future breeding applications.

Improvement of soil fertility and rice yield after long-term application of cow manure combined with inorganic fertilizers

Fertilization is an effective technique to improve soil fertility and increase crop yield.The long-term effects of different fertilizers on soil considerably vary.Over 38 consecutive years of different fertilization positioning experiments in a double cropping rice field of Qiyang Red Soil Experimental Station,seven different fertilization treatments including CK(no fertilization),NPK(nitrogen,phosphorus,and potassium fertilizer),M(cow manure),NPKM(nitrogen,phosphorus,and potassium with cow manure),NPM(nitrogen and phosphorus with cow manure),NKM(nitrogen and potassium with cow manure),and PKM(phosphorus and potassium with cow manure)were applied to study the effects on rice yield,soil fertility,and nutrient apparent balance in a paddy field.The results showed that the annual average yields of rice in NPKM,NPM,NKM,PKM,M,NPK and CK treatments ranged from 6214 to 11562 kg ha-1.Yields under long-term organic and inorganic treatments(NPKM,NPM,NKM and PKM)were 22.58,15.35,10.53 and 4.41%,respectively,greater than under the NPK treatment.Soil organic carbon(SOC),total nitrogen(TN),available nitrogen(AN)and available potassium(AK)concentration with long-term organic and inorganic treatment(NPKM,NPM,NKM and PKM)were significantly higher than in inorganic fertilizer(NPK)treatments.Soil total phosphorus(TP)and available phosphorus(AP)contentration with organic fertilizer combined with inorganic N and P fertilizer treatment(NPKM,NPM and PKM)were significantly higher than with inorganic fertilizer alone(NPK treatments).The average annual rice yield(11562 kg ha-1),SOC(20.88 g kg-1),TN(2.30 g kg-1),TP(0.95 g kg-1),TK(22.50 g kg-1)and AP(38.94 mg kg-1)concentrations were the highest in the NPKM treatment.The soil AN concentration(152.40 mg kg-1)and AK contentration(151.00 mg kg-1)were the highest in the NKM treatment.N and P application led to a surplus of nitrogen and phosphorus in the soil,but NPKM treatment effectively reduced the surplus compared with other treatments.Soils under all treatments were deficient in potassium.Correlation analysis showed that SOC,TN,AN,TP,and AP contentration was significantly correlated with rice yield;the correlation coefficients were 0.428,0.496,0.518,0.501,and 0.438,respectively.This study showed that the combined application of N,P,and K with cow manure had important effects on rice yield and soil fertility,but balanced application of N,P,and K with cow manure was required.

Genetic diversity,population structure and core collection analysis of Hunan tea plant germplasm through genotyping-by-sequencing

Tea is considered to be a well-known and widely consumed beverage and Hunan province is rich in tea plant germplasm.In order to better conserve and utilize Hunan tea plant resources,110 tea accessions from seven geographical origins were used to assess genetic diversity of Hunan tea plant germplasm through genotyping by sequencing(GBS)technology.As a result,a total of 311,044 high-quality single nucleotide polymorphism(SNP)markers were obtained.Population structure,phylogenetic relationships and principal component analysis(PCA)divided the entire accessions into three groups.The genetic diversity and population differentiation analysis showed that the mean observed heterozygosity(Ho)ranged from 0.16 to 0.24,while the mean polymorphic information content(PIC)ranged from 0.14 to 0.17,and mean minor allele frequency(MAF)ranged from 0.11 to 0.14.Analysis of molecular variance(AMOVA)indicated that 81.38%of the total variance was derived from within populations,which suggested a rich genetic diversity in Hunan tea germplasms.Furthermore,a core tea germplasm set was developed,which was comprised of 22 tea plant accessions and maintained the whole genetic diversity of the entire collection.This work should be valuable for conservation and utilization of tea germplasm in Hunan。

Technical Regulations for Under-forest Cultivation of Paris polyphylla Smith in Hunan Province

In this paper,the seedling raising,cultivation,field management,pest and disease control,harvesting and processing of Paris polyphylla Smith under the forest in Hunan Province were scientifically standardized,to provide technical support for standardized cultivation and sustainable development of Paris polyphylla Smith industry in Hunan Province.

The effects of co-utilizing green manure and rice straw on soil aggregates and soil carbon stability in a paddy soil in southern China

The co-utilization of green manure (GM) and rice straw (RS) in paddy fields has been widely applied as an effective practice in southern China.However,its effects on soil aggregate and soil organic carbon (SOC) stability remain unclear.In the present study,the effect of GM,RS,and co-utilization of GM and RS on particle size distribution of soil aggregates and SOC density fractions were measured in a field experiment.The experiment included six treatments,i.e.,winter fallow (WF) without RS return (Ctrl),WF with 50%RS return (1/2RS),WF with 100%RS return (RS),GM without RS return (GM),GM with 50%RS return (GM1/2RS) and GM with 100%RS return (GMRS).The results showed that the proportion of small macro-aggregates (0.25–2 mm) and the mean weight diameter (MWD) of aggregates in the GMRS treatment was greater (by 18.9 and 3.41%,respectively) than in the RS treatment,while the proportion of silt+clay particles (<0.053 mm) was lower (by 14.4%).The concentration of SOC in microaggregates (0.053–0.25 mm)and silt+clay particles was higher in the GMRS treatment than in GM and RS treatments individually.The concentration and proportion of free light organic carbon (fLOC) in aggregates of various particle sizes and bulk soil was greater in the GMRS treatment than the RS treatment,whereas the concentration and proportion of mineral-associated organic carbon in small macroaggregates,microaggregates,and bulk was lower in the GMRS treatment than in the RS treatment.The proportion of intra-aggregate particulate organic carbon (iPOC) was greater in the GMRS treatment than in GM treatment.The GMRS treatment had strong positive effects on iPOC in small macroaggregates,suggesting that SOC was transferred from fLOC to iPOC.In conclusion,co-utilizing green manure and rice straw cultivated the SOC pool by increasing the concentration of fLOC and improved soil carbon stability by promoting the sequestration of organic carbon in iPOC as a form of physical protection.

Safety evaluation and whole genome sequencing for revealing the ability of Penicillium oxalicum WX-209 to safely and effectively degrade citrus segments

The microbial potential of Penicillium has received critical attention.The present research aimed to elucidate the efficacy of crude enzyme secreted from Penicillium oxalicum WX-209 in degrading citrus segments and evaluate the safety of the process.Results showed that citrus segment membranes gradually dissolved after treatment with the crude enzyme solution,indicating good degradation capability.No significant differences in body weight,food ingestion rate,hematology,blood biochemistry,and weight changes of different organs were found between the enzyme intake and control groups.Serial experiments showed that the crude enzyme had high biological safety.Moreover,the whole genome of P.oxalicum WX-209 was sequenced by PacBio and Illumina platforms.Twenty-five scaffolds were assembled to generate 36 Mbp size of genome sequence comprising 11369 predicted genes modeled with a GC content of 48.33%.A total of 592 genes were annotated to encode enzymes related to carbohydrates,and some degradation enzyme genes were identified in strain P.oxalicum WX-209.

The 218th amino acid change of Ser to Ala in TaAGPS-7A increases enzyme activity and grain weight in bread wheat

ADP-glucose pyrophosphorylase(AGPase)influences cereal productivity.There are few reports on the function of cytosolic AGPase small subunit in bread wheat(TaAGPS).In the present study,TaAGPS was preferentially expressed in developing endosperm during grain-filling stages in bread wheat.TaAGPS allelic variations were characterized in 143 wheat accessions by PacBio RS II sequencing.Two haplotypes(TaAGPS-7A-TG and TaAGPS-7A-CT)of TaAGPS-7A were identified and corresponding functional markers were developed,whereas no variants of TaAGPS-7B and TaAGPS-7D were detected.TaAGPS-7A was associated with thousand-kernel weight(TKW)by haplotype–trait association analysis in two populations.Near-isogenic lines(NILs)with TaAGPS-7A-TG showed higher TKW and total kernel starch content than those with TaAGPS-7A-CT,owing to the higher AGPase activity of TaAGPS-7A-TG than TaAGPS-7A-CT both in vitro and in vivo.Overexpression of TaAGPS-7A-TG in bread wheat doubled the transcription levels of TaAGPS and increased AGPase activity by 55.7%,resulting in a 3.0-g higher TKW than in the wild type(WT).Knockdown of TaAGPS led to reduced expression of TaAGPS,AGPase activity,and TKW than in the WT.Thus,owing to the 218th amino acid change of Ser to Ala in TaAGPS-7A,the favorable haplotype TaAGPS-7A-TG showed higher AGPase activity,resulting in higher kernel starch content and grain weight.This finding could be applied to increasing starch content and grain weight in bread wheat.

Development and Application of a Novel Functional Marker for Fragrance in Rice

Fragrant rice is favored worldwide by consumers and rice breeders,due to its full aroma and high nutritional value.Loss of function of the betaine aldehyde dehydrogenase gene(OsBADH2)leads to aroma in rice.Our previous sequencing results showed that fragrant rice Chuan 7 belongs to badh2-E14 type,with a 1 bp insertion mutation in exon 14(Sun et al,2021),and this allele mutation is mainly distributed in Nepal and India(Kovach et al,2009).

A bacterial protein Rhp-PSP inhibits plant viral proliferation through endoribonuclease activity

Plant virus causes massive crop losses globally.However,there is currently no effective measure to control plant viral disease.Previously,we identify an antiviral protein Rhp-PSP,produced by the bacterial Rhodopseudomonas palustris strain JSC-3b.In this study,we discover that the antiviral activity of Rhp-PSP relies on its endoribonuclease activity.Converting the arginine(R)residue at position 129 onto alanine(A)abolishs its endoribonuclease activity on coat protein(CP)RNA of tobacco mosaic virus(TMV),consequentially,compromises the antiviral activity of Rhp-PSP.Further investigation demonstrates that,the mutant Rhp-PSP^(R129A)is unable to form the homotrimer as the wild type,indicating the importance of quaternary junction for the endoribonuclease activity.Overexpression of Rhp-PSP in Nicotiana benthamiana significantly enhances the resistance against TMV of seedlings,while expression of Rhp-PSP^(R129A)did not,confirming that endoribonuclease activity is responsible for the antiviral activity of Rhp-PSP.In addition,foliar spray of Rhp-PSP solution on tomato and pepper plants significantly reduces the disease index of viral diseases,indicating that Rhp-PSP shows potential to develop antiviral agent in practice.

CsABF3-activated CsSUT1 pathway is implicated in pre-harvest water deficit inducing sucrose accumulation in citrus fruit

Pre-harvest water deficit(PHWD)plays an important role in sugar accumulation of citrus fruit.However,the mechanism is not known well.Here,it was confirmed that PHWD promoted sucrose accumulation of citrus fruit,but had limited effect on fructose,glucose and total acid.A sucrose transporter,Cs SUT1,which localizes to the plasma membrane,was demonstrated to function in sucrose transport induced by PHWD.Compared to wild-type,Cs SUT1 overexpression in citrus calli stimulated sucrose,fructose and glucose accumulation,while its silencing in juice sacs reduced sucrose accumulation.Increased sugar accumulation in transgenic lines enhanced plant drought tolerance,and resulted in decreased electrolyte leakage,malondialdehyde and hydrogen peroxide contents,as well as increased superoxide dismutase activity and proline contents.An abscisic acid(ABA)-responsive transcription factor,Cs ABF3,was found with a same expression pattern with Cs SUT1 under PHWD.Yeast one-hybrid,electrophoretic mobility shift assay and dual-luciferase assays all revealed that Cs ABF3 directly bound with the Cs SUT1 promoter by ABA responsive elements.When Cs ABF3 was overexpressed in citrus calli,the sucrose,fructose and glucose concentration increased correspondingly.Further,transgenic studies demonstrated that Cs ABF3 could affect sucrose accumulation by regulating Cs SUT1.Overall,this study revealed a regulation of Cs ABF3 promoting Cs SUT1 expression and sucrose accumulation in response to PHWD.Our results provide a detail insight into the quality formation of citrus fruit.

Seed Storability in Rice: Physiological Foundations, Molecular Mechanisms, and Applications in Breeding

Long-term storage of crop seeds is critical for the conservation of germplasm resources, ensuring food supply, and supporting sustainable production. Rice, as a major food staple, has a substantial stock for consumption and production worldwide. However, its food value and seed viability tend to decline during storage. Understanding the physiological responses and molecular mechanisms of aging tolerance forms the basis for enhancing seed storability in rice. This review outlines the latest progress in influential factors, evaluation methods, and identification indices of seed storability. It also discusses the physiological consequences, molecular mechanisms, and strategies for breeding aging-tolerant rice in detail. Finally, it highlights challenges in seed storability research that require future attention. This review offers a theoretical foundation and research direction for uncovering the mechanisms behind seed storability and breeding aging-tolerant rice.

Single-cell transcriptome sequencing reveals the mechanism regulating rice plumule development

Seed plumules comprise multiple developing tissues and are key sites for above-ground plant organ morphogenesis.Here,the spatial expression of genes in developing rice seed plumules was characterized by single-cell transcriptome sequencing in Zhongjiazao 17,a popular Chinese indica rice cultivar.Of 15 cell clusters,13 were assigned to cell types using marker genes and cluster-specific genes.Marker genes of multiple cell types were expressed in several clusters,suggesting a complex developmental system.Some genes for signaling by phytohormones such as abscisic acid were highly expressed in specific clusters.Various cis-elements in the promoters of genes specifically expressed in cell clusters were calculated,and some key hormone-related motifs were frequent in certain clusters.Spatial expression patterns of genes involved in rapid seed germination,seedling growth,and development were identified.These findings enhanced our understanding of cellular diversity and specialization within plumules of rice,a monocotyledonous model crop.

Effects of Long-Term Winter Planted Green Manure on Distribution and Storage of Organic Carbon and Nitrogen in Water-Stable Aggregates of Reddish Paddy Soil Under a Double-Rice Cropping System

In agricultural systems, maintenance of soil organic matter has long been recognized as a strategy to reduce soil degradation. Manure amendments and green manures are management practices that can increase some nutrient contents and improve soil aggregation. We investigated the effects of 28 yr of winter planted green manure on soil aggregate-size distribution and aggregateassociated carbon（C） and nitrogen（N）. The study was a randomized completed block design with three replicates. The treatments included rice-rice-fallow, rice-rice-rape, rice-rice-Chinese milk vetch and rice-rice-ryegrass. The experiment was established in 1982 on a silty light clayey paddy soil derived from Quaternary red clay（classified as Fe-Accumuli-Stagnic Anthrosols） with continuous early and late rice. In 2009, soil samples were collected（0-15 cm depth） from the field treatment plots and separated into water-stable aggregates of different sizes（i.e., 〉5, 2-5, 1-2, 0.5-1, 0.25-0.5 and 〈0.25 mm） by wet sieving. The long-term winter planted green manure significantly increased total C and N, and the formation of the 2-5-mm water-stable aggregate fraction. Compared with rice-rice-rape, rice-rice-Chinese milk vetch and rice-rice-ryegrass, the rice-rice-fallow significantly reduced 2-5-mm water-stable aggregates, with a significant redistribution of aggregates into micro-aggregates. Long-term winter planted green manure obviously improved C/N ratio and macro-aggregate-associated C and N. The highest contribution to soil fertility was from macro-aggregates of 2-5 mm in most cases.

Long-Term Effect of Fertilizer and Rice Straw on Mineral Composition and Potassium Adsorption in a Reddish Paddy Soil

Increasing K＋ adsorption can be an effective alternative in building an available K pool in soils to optimize crop recovery and minimize losses into the environment. We hypothesized that long-term fertilization might change K＋ adsorption because of changes in the chemical and mineralogical properties of a rice （Oryza sativa L.）. The aims of this study were （i） to determine clay minerals in paddy soil clay size fractions using X-ray diffraction methods and a numerical diagramdecomposition method; （ii） to measure K＋ adsorption isotherms before and after H202 oxidation of organic matter, and （iii） to investigate whether K＋ adsorption is correlated with changes in soil chemical and mineral properties. The 30-yr longterm fertilization treatments caused little change in soil organic C （SOC） but a large variation in soil mineral composition. The whole-clay fraction （〈5 Jam） corresponded more to the fertilization treatment than the fine-clay fraction （〈1 gin） in terms of percentage of illite peak area. The total percentage of vermiculite-chlorite peak area was significantly negatively correlated with the total percentage ofillite peak area in the 〈5 lam soil particles （R=-0.946, P〈0.0006）. Different fertilization treatments gave significantly different results in K＋ adsorption. The SOC oxidation test showed positive effects of SOC on K＋ adsorption at lower K＋ concentration （≤120 mg L-0 and negative effects at higher K＋ concentration （240 mg L-l）. The K＋ adsorption by soil clay minerals after SOC oxidization accounted for 60-158% of that by unoxidized soils, suggesting a more important role of soil minerals than SOC on K＋ adsorption. The K＋ adsorption potential was significantly correlated to the amount of poorly crystallized illite present （R--0.879, P=0.012）. The availability of adsorbed K＋ for plant growth needs further study.

Effects of Subsoiling on Soil Moisture Under No-Tillage for Two Years

In order to improve the water use efficiency under conservation tillage, the effects of subsoiling on soil moisture under notillage were studied. An experiment of 40 cm subsoiling in a field kept under no-tillage for 2 years was operated from 2005 to 2006. Based on the data of the soil moisture and crop yield, the physical basis of subsoiling for water conservation and yield increase was analyzed. The results showed that the soil water storage under subsoiling, from the soil surface to a depth of 100 cm was more than that under no-tillage for the growth season. In the 0-100 cm soil depth, the soil moisture in 50-100 cm depth under subsoiling was more compared with no-tillage, which increased when it＇s drought and decreased when it＇s rainy with the increase in soil depth. Compared with no-tillage, subsoiling could reduce the water consumption of oats in the 0-50 cm depth and increase the water consumption in the 50-100 cm depth. Also, subsoiling increased the yield by 18.29% and the water use efficiency by 16.8% in a two-year average. The effects of subsoiling on water conservation and yield increase were affected by precipitation, and a well-proportioned rainfall was better to increase yield and water use efficiency. Meanwhile, subsoiling decreased bulk density, which increased with the available precipitation. Subsoiling under no-tillage is the effective rotation tillage to contain more soil moisture and improve water use efficiency in ecotone of North China.

Identification of QTLs and Validation of qCd-2 Associated with Grain Cadmium Concentrations in Rice

Cadmium(Cd) is one of heavy metals harmful to human health. As rice is the main staple food in Asia and Cd is easily contaminated in rice, the molecular regulation of Cd accumulation should be explored. In this study, a recombinant inbred population derived from Xiang 743/Katy was grown in Cd-polluted fields and used to map the quantitative trait loci(QTLs) for Cd accumulation in rice grains. We identified seven QTLs distributed on chromosomes 2, 3, 6, 7, 8 and 10. These QTLs displayed phenotypic variances of 58.50% and 40.59% in 2014 and 2015, respectively. Two QTLs, qCd-2 and qCd-7, were identified in both the two years. qCd-2 was detected on the interval of RM250–RM207 on chromosome 2, with an LOD of 2.51 and a phenotypic contribution of 13.75% in 2014, and an LOD of 3.35 and a phenotypic contribution of 14.16% in 2015. qCd-7 co-localized with the cloned qCdT7 on chromosome 7 and may represent the correct candidate. The other five QTLs were detected only in one year. To further confirm the effects of qCd-2, a residual heterozygous line designated as RHL945, with a heterozygous interval of RM263–RM207 on chromosome 2, was selected from the recombinant inbred population and used to develop an F2 population consisting of 155 individual plants. By incorporating further simple sequence repeat markers into the segmental linkage map of the target region, qCd-2 was delimited in the interval of RM5404–RM3774, with an LOD value of 4.38 and a phenotypic contribution of 15.52%. These results reflected the genetic regulation of grain Cd in rice and paved the way for the future cloning of qCd-2.

Substitution of chemical fertilizer by Chinese milk vetch improves the sustainability of yield and accumulation of soil organic carbon in a double-rice cropping system

The double-rice cropping system is a very important intensive cropping system for food security in China. There have been few studies of the sustainability of yield and accumulation of soil organic carbon (SOC) in the double-rice cropping system following a partial substitution of chemical fertilizer by Chinese milk vetch (Mv). We conducted a 10-year (2008–2017) field experiment in Nan County, South-Central China, to examine the double-rice productivity and SOC accumulation in a paddy soil in response to different fertilization levels and Mv application (22.5 Mg ha^–1). Fertilizer and Mv were applied both individually and in combination (sole chemical fertilizers, Mv plus 100, 80, 60, 40, and 0% of the recommended dose of chemical fertilizers, labeled as F100, MF100, MF80, MF60, MF40, and MF0, respectively). It was found that the grain yields of double-rice crop in treatments receiving Mv were reduced when the dose of chemical fertilizer was reduced, while the change in SOC stock displayed a double peak curve. The MF100 produced the highest double-rice yield and SOC stock, with the value higher by 13.5 and 26.8% than that in the F100. However, the grain yields increased in the MF80 (by 8.4% compared to the F100), while the SOC stock only increased by 8.4%. Analogous to the change of grain yield, the sustainable yield index (SYI) of double rice were improved significantly in the MF100 and MF80 compared to the F100, while there was a slight increase in the MF60 and MF40. After a certain amount of Mv input (22.5 Mg ha^–1), the carbon sequestration rate was affected by the nutrient input due to the stimulation of microbial biomass. Compared with the MF0, the MF100 and MF40 resulted in a dramatically higher carbon sequestration rate (with the value higher by 71.6 and 70.1%), whereas the MF80 induced a lower carbon sequestration rate with the value lower by 70.1% compared to the MF0. Based on the above results we suggested that Mv could partially replace chemical fertilizers (e.g., 40–60%) to improve or maintain the productivity and sustainability of the double-rice cropping system in South-Central China.

Spatio-temporal changes in rice area at the northern limits of the rice cropping system in China from 1984 to 2013

Rice area has been expanding rapidly during the past 30 years under the influence of global change in northeastern China, which is the northernmost region of rice cultivation in China. However, the spatio-temporal dynamic changes in rice area are still unclear, although they may have important policy implications for environmental protection and adaptation to climate change. In this study, we aimed to identify the dynamic changes of the rice area in Heilongjiang Province of northeastern China by extracting data from multiple Landsat images. The study used ground quadrats selected from Google Earth and the extraction of a confusion matrix to verify the accuracy of extraction. The overall accuracy of the extracted rice area was higher than 95% as a result of using the artificial neural network （ANN） classification method. The results showed that the rice area increased by approximately 2.4×10^6 ha during the past 30 years at an annual rate of 8.0×10^4 ha, and most of the increase occurred after 2000. The central latitude of the rice area shifted northwards from 46 to 47°N during the study period, and moved eastwards from 130 to 133°E. The rice expansion area accounted for 98% of the total change in rice area, and rice loss was notably rare. The rice expansion was primarily from dryland. In addition, rice cultivation in marshland and grassland played a minor role in the rice expansion in this region.