Optimizing the Bacillus thuringiensis(Bt)protein concentration in cotton:Coordinated application of exogenous amino acids and EDTA to reduce spatiotemporal variability in boll and leaf toxins

During the boll formation stage,cotton bolls exhibit the lowest expression of Bacillus thuringiensis(Bt)insecticidal proteins.Resistance to insects varies notably among different organs,which poses challenges for controlling cotton bollworms.Consequently,an experimental strategy was designed in the 2020-2021 cotton growing season to coordinate the enhancement of protein synthesis and the attenuation of degradation.Two Bt cultivars of Gossypium hirsutum,namely the hybrid Sikang 3 and the conventional Sikang 1,were used as test materials.Three treatments were applied at the peak flowering period:CK(the control),T1(amino acids),and T2(amino acids and EDTA).The results show that,in comparison to the CK group,the Bt protein contents were significantly increased in both cotton bolls and their subtending leaves under the T1 and T2 treatments.The maximum levels of increase observed were 67.5%in cotton bolls and 21.7%in leaves.Moreover,the disparity in Bt protein content between cotton bolls and their subtending leaves notably decreased by 31.2%.Correlation analysis suggested that the primary physiological mechanisms for augmenting Bt protein content involve increased protein synthesis and reduced protein catabolism,which are independent of Bt gene expression levels.Stepwise regression and path analysis revealed that elevating the soluble protein content and transaminase activity,while reducing the catabolic enzyme activities,are instrumental in enhancing the Bt protein content.Consequently,the coordinated application of amino acids and EDTA emerges as a strategy that can improve the overall resistance of Bt cotton and mitigate the spatiotemporal variations in Bt toxin concentrations in both cotton bolls and leaves.

Dynamics and genetic regulation of macronutrient concentrations during grain development in maize

Nitrogen(N), phosphorus(P), and potassium(K) are essential macronutrients that are crucial not only for maize growth and development, but also for crop yield and quality. The genetic basis of macronutrient dynamics and accumulation during grain filling in maize remains largely unknown. In this study, we evaluated grain N, P, and K concentrations in 206 recombinant inbred lines generated from a cross of DH1M and T877 at six time points after pollination. We then calculated conditional phenotypic values at different time intervals to explore the dynamic characteristics of the N, P, and K concentrations. Abundant phenotypic variations were observed in the concentrations and net changes of these nutrients. Unconditional quantitative trait locus(QTL) mapping revealed 41 non-redundant QTLs, including 17, 16, and 14 for the N, P, and K concentrations, respectively. Conditional QTL mapping uncovered 39 non-redundant QTLs related to net changes in the N, P, and K concentrations. By combining QTL, gene expression, co-expression analysis, and comparative genomic data, we identified 44, 36, and 44 candidate genes for the N, P, and K concentrations, respectively, including GRMZM2G371058 encoding a Doftype zinc finger DNA-binding family protein, which was associated with the N concentration, and GRMZM2G113967encoding a CBL-interacting protein kinase, which was related to the K concentration. The results deepen our understanding of the genetic factors controlling N, P, and K accumulation during maize grain development and provide valuable genes for the genetic improvement of nutrient concentrations in maize.

Grain Yield,Biomass Accumulation,and Leaf Photosynthetic Characteristics of Rice under Combined Salinity-Drought Stress

Simultaneous stresses of salinity and drought often coincide during rice-growing seasons in saline lands,primarily due to insufficient water resources and inadequate irrigation facilities.Consequently,combined salinity-drought stress poses a major threat to rice production.In this study,two salinity levels(NS,non-salinity;HS,high salinity)along with three drought treatments(CC,control condition;DJ,drought stress imposed at jointing;DH,drought stress imposed at heading)were performed to investigate their combined influences on leaf photosynthetic characteristics,biomass accumulation,and rice yield formation.Salinity,drought,and their combination led to a shortened growth period from heading to maturity,resulting in a reduced overall growth duration.Grain yield was reduced under both salinity and drought stress,with a more substantial reduction under the combined salinity-drought stress.The combined stress imposed at heading caused greater yield losses in rice compared with the stress imposed at jointing.Additionally,the combined salinity-drought stress induced greater decreases in shoot biomass accumulation from heading to maturity,as well as in shoot biomass and nonstructural carbohydrate(NSC)content in the stem at heading and maturity.However,it increased the harvest index and NSC remobilization reserve.Salinity and drought reduced the leaf area index and SPAD value of flag leaves and weakened the leaf photosynthetic characteristics as indicated by lower photosynthetic rates,transpiration rates,and stomatal conductance.These reductions were more pronounced under the combined stress.Salinity,drought,and especially their combination,decreased the activities of ascorbate peroxidase,catalase,and superoxide dismutase,while increasing the contents of malondialdehyde,hydrogen peroxide,and superoxide radical.Our results indicated a more significant yield loss in rice when subjected to combined salinity-drought stress.The individual and combined stresses of salinity and drought diminished antioxidant enzyme activities,inhibited leaf photosynthetic functions,accelerated leaf senescence,and subsequently lowered assimilate accumulation and grain yield.

Mepiquat chloride increases the Cry1Ac protein content of Bt cotton under high temperature and drought stress by regulating carbon and amino acid metabolism

The effects of mepiquat chloride(DPC)on the Cry1Ac protein content in Bacillus thuringiensis(Bt)cotton boll shells under high temperature and drought stress were investigated to provide a theoretical reference for Bt cotton breeding and high-yield and-efficiency cotton cultivation.This study was conducted using Bt cotton cultivar‘Sikang 3'during the 2020 and 2021 growing seasons at Yangzhou University Farm,Yangzhou,Jiangsu Province,China.Potted cotton plants were exposed to high temperature and drought stress,and sprayed with either 20 mg L^(-1)DPC or water(CK).Seven days after treatment,the Cry1Ac protein content,α-ketoglutarate content,pyruvic acid content,glutamate synthase activity,glutamic oxaloacetic transaminase activity,soluble protein content,and amino acid content were measured,and transcriptome sequencing was performed.DESeq was used for differential gene analysis.Under the DPC treatment,the Cry1Ac protein content increased by 4.7-11.9% compared to CK.Theα-ketoglutarate content,pyruvic acid content,glutamate synthase activity,glutamic oxaloacetic transaminase activity,soluble protein content,and amino acid content all increased.Transcriptome analysis revealed 7,542 upregulated genes and 10,449 downregulated genes for DPC vs.CK.Gene ontology(GO)and Kyoto Encyclopedia of Gene and Genomes(KEGG)analyses showed that the differentially expressed genes were mainly involved in biological processes,such as carbon and amino acid metabolism.For example,genes encoding 6-phosphofructokinase,pyruvate kinase,glutamic pyruvate transaminase,pyruvate dehydrogenase,citrate synthase,isocitrate dehydrogenase,2-oxoglutarate dehydrogenase,glutamate synthase,1-pyrroline-5-carboxylate dehydrogenase,glutamic oxaloacetic transaminase,amino-acid N-acetyltransferase,and acetylornithine deacetylase were all significantly upregulated.The DPC treatment increased pyruvate,α-ketoglutarate,and oxaloacetate by increasing the operational rate of the glycolytic pathway of the citric acid cycle.It also significantly upregulated the genes encoding glutamate synthase,pyrrolidine-5-carboxylic acid dehydrogenase,glutamate oxaloacetate transaminase,and N-acetylglutamate synthetase,while it downregulated the genes encoding glutamine synthetase.Therefore,the synthesis of aspartic acid,glutamic acid,pyruvate,and arginine increased after treatment with DPC,and the Cry1Ac protein content was increased by regulating carbon and amino acid metabolism.

The response of roots and the rhizosphere environment to integrative cultivation practices in paddy rice

Integrative cultivation practices(ICPs)are essential for enhancing cereal yield and resource use efficiency.However,the effects of ICP on the rhizosphere environment and roots of paddy rice are still poorly understood.In this study,four rice varieties were produced in the field.Each variety was treated with six different cultivation techniques,including zero nitrogen application(0 N),local farmers’practice(LFP),nitrogen reduction(NR),and three progressive ICP techniques comprised of enhanced fertilizer N practice and increased plant density(ICP1),a treatment similar to ICP1 but with alternate wetting and moderate drying instead of continuous flooding(ICP2),and the same practices as ICP2 with the application of organic fertilizer(ICP3).The ICPs had greater grain production and nitrogen use efficiency than the other three methods.Root length,dry weight,root diameter,activity of root oxidation,root bleeding rate,zeatin and zeatin riboside compositions,and total organic acids in root exudates were elevated with the introduction of the successive cultivation practices.ICPs enhanced nitrate nitrogen,the activities of urease and invertase,and the diversity of microbes(bacteria)in rhizosphere and non-rhizosphere soil,while reducing the ammonium nitrogen content.The nutrient contents(ammonium nitrogen,total nitrogen,total potassium,total phosphorus,nitrate,and available phosphorus)and urease activity in rhizosphere soil were reduced in all treatments in comparison with the non-rhizosphere soil,but the invertase activity and bacterial diversity were greater.The main root morphology and physiology,and the ammonium nitrogen contents in rhizosphere soil at the primary stages were closely correlated with grain yield and internal nitrogen use efficiency.These findings suggest that the coordinated enhancement of the root system and the environment of the rhizosphere under integrative cultivation approaches may lead to higher rice production.

Optimizing nitrogen management can improve stem lodging resistance and stabilize the grain yield of japonica rice in rice-crayfish coculture systems

Nitrogen(N)significantly affects rice yield and lodging resistance.Previous studies have primarily investigated the impact of N management on rice lodging in conventional rice monoculture(RM);however,few studies have performed such investigations in rice-crayfish coculture(RC).We hypothesized that RC would increase rice lodging risk and that optimizing N application practices would improve rice lodging resistance without affecting food security.We conducted a two-factor(rice farming mode and N management practice)field experiment from2021 to 2022 to test our hypothesis.The rice farming modes included RM and RC,and the N management practices included no nitrogen fertilizer,conventional N application,and optimized N treatment.The rice yield and lodging resistance characteristics,such as morphology,mechanical and chemical characteristics,anatomic structure,and gene expression levels,were analyzed and compared among the treatments.Under the same N application practice,RC decreased the rice yield by 11.1-24.4% and increased the lodging index by 19.6-45.6% compared with the values yielded in RM.In RC,optimized N application decreased the plant height,panicle neck node height,center of gravity height,bending stress,and lodging index by 4.0-4.8%,5.2-7.8%,0.5-4.5%,5.5-10.5%,and 1.8-19.5%,respectively,compared with those in the conventional N application practice.Furthermore,it increased the culm diameter,culm wall thickness,breaking strength,and non-structural and structural carbohydrate content by 0.8-4.9%,2.2-53.1%,13.5-19.2%,2.2-24.7%,and 31.3-87.2%,respectively.Optimized N application increased sclerenchymal and parenchymal tissue areas of the vascular bundle at the culm wall of the base second internode.Furthermore,optimized N application upregulated genes involved in lignin and cellulose synthesis,thereby promoting lower internodes on the rice stem and enhancing lodging resistance.Optimized N application in RC significantly reduced the lodging index by 1.8-19.5%and stabilized the rice yield(>8,570 kg ha~(-1)on average).This study systematically analyzed and compared the differences in lodging characteristics between RM and RC.The findings will aid in the development of more efficient practices for RC that will reduce N fertilizer application.

Differing responses of root morphology and physiology to nitrogen application rates and their relationships with grain yield in rice

Root morphology and physiology influence aboveground growth and yield formation in rice.However,root morphological and physiological differences among rice varieties with differing nitrogen(N)sensitivities and their relationship with grain yield are still unclear.In this study,rice varieties differing in N sensitivity over many years of experiments were used.A field experiment with multiple N rates(0,90,180,270,and 360 kg ha^(-1))was conducted to elucidate the effects of N application on root morphology,root physiology,and grain yield.A pot experiment with root excision and exogenous application of 6-benzyladenine(6-BA)at heading stage was used to further verify the above effects.The findings revealed that(1)under the same N application rate,N-insensitive varieties(NIV)had relatively large root biomass(root dry weight,length,and number).Grain yield was associated with root biomass in NIV.The oxidation activity and zeatin(Z)+zeatin riboside(ZR)contents in roots obviously and positively correlated with grain yield in N-sensitive varieties(NSV),and accounted for its higher grain yield than that of NIV at lower N application rates(90 and 180 kg ha^(-1)).(2)The root dry weight required for equal grain yield of NIV was greater than that of NSV.Excision of 1/10 and 1/8 of roots at heading stage had no discernible effect on the yield of Liangyoupeijiu(NIV),and it significantly reduced yield by 11.5%and 21.3%in Tianyouhuazhan(NSV),respectively,compared to the treatment without root excision.The decrease of filled kernels and grain weight after root excision was the primary cause for the yield reduction.Root excision and exogenous 6-BA application after root excision had little influence on the root activity of NIV.The oxidation activity and Z+ZR contents in roots of NSV decreased under root excision,and the increase in the proportion of excised roots aggravated these effects.The application of exogenous 6-BA increased the root activity of NSV and increased filled kernels and grain weight,thereby reducing yield loss after root excision.Thus,the root biomass of NIV was large,and there may be a phenomenon of"root growth redundancy."Vigorous root activity was an essential feature of NSV.Selecting rice varieties with high root activity or increasing root activity by cultivation measures could lead to higher grain yield under lower N application rates.

Translocation and Distribution of Carbon-Nitrogen in Relation to Rice Yield and Grain Quality as Affected by High Temperature at Early Panicle Initiation Stage

Due to climate change, extreme heat stress events have become more frequent, adversely affecting rice yield and grain quality. The accumulation and translocation of dry matter and nitrogen substances are essential for rice yield and grain quality. To assess the impact of high temperature stress(HTS) at the early panicle initiation(EPI) stage on the accumulation, transportation, and distribution of dry matter and nitrogen substances in various organs of rice, as well as the resulting effects on rice yield and grain quality, pot experiments were conducted using an indica rice cultivar Yangdao 6(YD6) and a japonica rice cultivar Jinxiangyu 1(JXY1) under both normal temperature(32 ℃/26 ℃) and high temperature(38 ℃/29 ℃) conditions. The results indicated that exposure to HTS at the EPI stage significantly decreased rice yield by reducing spikelet number per panicle, grain-filling rate, and grain weight. However, it improved the nutritional quality of rice grains by increasing protein and amylose contents. The reduction in nitrogen and dry matter accumulation accounted for the changes in spikelet number per panicle, grain-filling rate, and grain size. Under HTS, the decrease in nitrogen accumulation accompanied by the reduction in dry matter may be due to the down-regulation of leaf net photosynthesis and senescence, as evidenced by the decrease in nitrogen content. Furthermore, the decrease in sink size limited the translocation of dry matter and nitrogen substances to grains, which was closely related to the reduction in grain weight and the deterioration of grain quality. These findings significantly contribute to our understanding of the mechanisms of HTS on grain yield and quality formation from the perspective of dry matter and nitrogen accumulation and translocation. Further efforts are needed to improve the adaptability of rice varieties to climate change in the near future.

Enhancing boll protein synthesis and carbohydrate conversion by the application of exogenous amino acids at the peak flowering stage increased the boll Bt toxin concentration and lint yield in cotton

In Bacillus thuringenesis(Bt) transgenic cotton, the cotton boll has the lowest insecticidal protein content when compared to the other organs. The present study investigated the effects of amino acid spray application at the peak flowering stage on the cotton boll Bt toxin concentration and yield formation. Boll protein synthesis and carbohydrate conversion were also studied to reveal the fundamental mechanism. Three treatments(i.e., CK, the untreated control;LA1, five amino acids;LA2, 21 amino acids) were applied to two Bt cultivars of G. hirsutum(i.e., the hybrid Sikang 3 and the conventional Sikang 1) in the cotton-growing seasons during 2017 and 2018. Amino acid spray application at the peak flowering stage resulted in an increase of 5.2–16.4% in the boll Bt protein concentration and an increase of 5.5–11.3%in the seed cotton yield, but there was no difference between the two amino acid treatments. In addition, amino acid applications led to increases in the amino acid content, soluble protein content, glutamate pyruvate transaminase(GPT)activity, glutamate oxaloacetate transaminase(GOT) activity, glucose content, fructose content and soluble acid invertase(SAI) activity. This study also found that Bt protein content, enhanced boll number and the weight of opened bolls were closely related to carbon and nitrogen metabolism. The Bt protein content had significant linear positive correlations with amino acid and soluble protein contents. Enhanced boll number had significant linear positive correlations with the GPT and GOT activities from 15–25 days after flowering(DAF). The weight of opened bolls from 55–65 DAF had a significant linear positive correlation with the SAI activity. These results indicate that the enhancement of boll protein synthesis and carbohydrate conversion by amino acid application resulted in a simultaneous increase in the boll Bt protein concentration and cotton lint yield.

Grain yield,nitrogen use efficiency and physiological performance of indica/japonica hybrid rice in response to various nitrogen rates

Utilizing the heterosis of indica/japonica hybrid rice(IJHR)is an effective way to further increase rice grain yield.Rational application of nitrogen(N)fertilizer plays a very important role in using the heterosis of IJHR to achieve its great yield potential.However,the responses of the grain yield and N utilization of IJHR to N application rates and the underlying physiological mechanism remain elusive.The purpose of this study was to clarify these issues.Three rice cultivars currently used in rice production,an IJHR cultivar Yongyou 2640(YY2640),a japonica cultivar Lianjing 7(LJ-7)and an indica cultivar Yangdao 6(YD-6),were grown in the field with six N rates(0,100,200,300,400,and 500 kg ha^(-1))in 2018 and 2019.The results showed that with the increase in N application rates,the grain yield of each test cultivar increased at first and then decreased,and the highest grain yield was at the N rate of 400 kg ha^(-1)for YY2640,with a grain yield of 13.4 t ha^(-1),and at 300 kg ha^(-1)for LJ-7 and YD-6,with grain yields of 9.4–10.6 t ha^(-1).The grain yield and N use efficiency(NUE)of YY2640 were higher than those of LJ-7 or YD-6 at the same N rate,especially at the higher N rates.When compared with LJ-7 or YD-6,YY2640 exhibited better physiological traits,including greater root oxidation activity and leaf photosynthetic rate,higher cytokinin content in the roots and leaves,and more remobilization of assimilates from the stem to the grain during grain filling.The results suggest that IJHR could attain both higher grain yield and higher NUE than inbred rice at either low or high N application rates.Improved shoot and root traits of the IJHR contribute to its higher grain yield and NUE,and a higher content of cytokinins in the IJHR plants plays a vital role in their responses to N application rates and also benefits other physiological processes.

Effects of planting methods on yield and quality of different types of japonica rice in northern Jiangsu plain, China

Mechanical transplanting with carpet seedlings(MC) and mechanical direct seeding(MD) are newly developed planting methods, which increase in popularity and planted area each year. Knowing the difference for yield and rice quality under different planting methods is of great importance for the development of high quality and yield cultivation techniques under mechanical conditions. Therefore, three kinds of japonica rice including hybrid japonica rice, inbreed japonica rice, and soft rice were adopted as materials. And the differences in the quality of processing, appearance, cooking and eating quality, nutrition, and the rapid viscosity analyzer(RVA) profile were studied to reveal the effects of planting methods on yield and quality of different types of japonica rice. Results showed that the milled rice and head rice rates under MC was significantly higher than those under MD, and the processing quality of inbreed japonica rice was the most stable. Compared with MC, length/width ratio of rice under MD was significantly increased, and chalkiness rate, size, and degree were significantly decreased. The protein content under MD was lower than that under MC. MC showed higher peak viscosity and breakdown value than MD. The taste value was the greatest for soft rice, followed by inbreed japonica rice, and then by japonica hybrid rice, with no significant differences resulting from planting methods. Compared with MC, MD significantly improved the appearance quality, though processing quality and nutritional quality were decreased. And there was no significant difference in cooking and eating quality between MC and MD. Under different planting methods, the appearance quality of inbreed japonica rice changed the most and the processing quality was the most stable. The nutritional, cooking and eating quality of soft rice changed the least. Therefore, according to the different planting methods and market needs, selecting the appropriate rice varieties can reduce the risks in rice production and achieve good rice quality.

The effect of integrative crop management on root growth and methane emission of paddy rice

In previous studies, integrative crop management (ICM) improved shoot growth and grain yield of rice (Oryza sativa L.). However, little is known about the effect of ICM on root growth and methane (CH4) emission of paddy rice. In this study, two rice varieties, Wuyunjing 24 and Yongyou 2640, were grown. A field experiment was conducted with three crop management treatments including zero nitrogen fertilization (0N), local farmer practice (LFP), and ICM. Root morphophysiological traits and CH4 emission from the paddy field were investigated. ICM significantly increased mean grain yield by 29.9%, with the effect attributed mainly to an increase in mean total number of spikelets by 26.4% compared to LFP. ICM increased root and shoot biomass, root length, number of roots, root oxidation activity (ROA), root bleeding rate, and root total and active absorbing surface area by respectively 24.4%, 25.7%, 17.1%, 9.3%, 18.7%, 29.5%, 12.1%, and 24.7%. The concentrations of malic, succinic, and acetic acids in root exudates were respectively 5.8%, 6.0%, and 10.5% higher in ICM than in LFP. Compared to LFP, ICM significantly decreased the rate of CH4 emission during emission peak stages and reduced total CH4 emission by 17.1%. The root morphophysiological traits were positively and significantly correlated with grain yield, whereas root length, specific root length, ROA, and root total and active absorbing surface area were negatively and significantly correlated with total CH4 emission. These results suggest that ICM could achieve the dual goals of increasing grain yield and reducing the greenhouse gas effect by improving the root morphology and physiological traits of paddy rice.

Comparison of grain yield and quality of different types of japonica rice cultivars in the northern Jiangsu plain,China

In recent years,an increasing number of different types of japonica rice cultivars have been released in the southern rice region of China.The grain yield and quality of these new cultivars showed significant differences in large scale planting.However,the causes of the differences remain little known.Therefore,three typical types of japonica rice cultivars were used in this study to investigate their grain yield and quality.A scanning calorimeter(DSC),X-ray powder diffractometer(XRD),rapid viscosity analyzer(RVA)and taste analyzer were used to evaluate the cooking and eating properties.The results showed that the yield of non-soft hybrid japonica rice cultivars was significantly higher than that of non-soft inbred japonica rice cultivars and soft inbred japonica rice cultivars.Soft inbred japonica rice cultivars had a low amylose content and moderate protein content,which are the main reasons for the superior cooking and eating quality.In addition,the relative crystallinity of soft inbred japonica rice cultivars was significantly higher than that of non-soft inbred and non-soft hybrid japonica rice cultivars,which is considered the major factor resulting in higher transition temperature and gelatinization enthalpy(ΔH_(gel)).Non-soft hybrid japonica rice cultivars had a higher number of large starch granules than soft inbred and non-soft inbred japonica rice cultivars.The setback value(SB)and breakdown value(BD),indirectly reflecting the cooking and eating quality of the three types of japonica rice cultivars,also confirmed that soft inbred japonica rice cultivars with a low SB value and a high BD value had better palatability than the other two types.This study provides guidance for future plantation of different types of japonica rice cultivars in large rice-producing areas.

Inhibition of protein degradation increases the Bt protein concentration in Bt cotton

Bacillus thuringiensis(Bt)cotton production is challenged by two main problems,i.e.,the low concentration of Bt protein at the boll setting stage and the lowest insect resistance in bolls among all the cotton plant’s organs.Therefore,increasing the Bt protein concentration at the boll stage,especially in bolls,has become the main goal for increasing insect resistance in cotton.In this study,two protein degradation inhibitors(ethylene diamine tetra acetic acid(EDTA)and leupeptin)were sprayed on the bolls,subtending leaves,and whole cotton plants at the peak flowering stage of two Bt cultivars(medium maturation Sikang 1(SK1))and early maturation Zhongmian 425(ZM425)in 2019 and 2020.The Bt protein content and protein degradation metabolism were assessed.The results showed that the Bt protein concentrations were enhanced by 21.3 to 38.8%and 25.0 to 38.6%in the treated bolls of SK1 and ZM425 respectively,while they were decreased in the subtending leaves of these treated bolls.In the treated leaves,the Bt protein concentrations increased by 7.6 to 23.5%and 11.2 to 14.9%in SK1 and ZM425,respectively.The combined application of EDTA and leupeptin to the whole cotton plant increased the Bt protein concentrations in both bolls and subtending leaves.The Bt protein concentrations in bolls were higher,increasing by 22.5 to 31.0%and 19.6 to 32.5%for SK1 and ZM425,respectively.The organs treated with EDTA or/and leupeptin showed reduced free amino acid contents,protease and peptidase activities and significant enhancements in soluble protein contents.These results indicated that inhibiting protein degradation could improve the protein content,thus increasing the Bt protein concentrations in the bolls or/and leaves of cotton plants.Therefore,the increase in the Bt protein concentration without yield reduction suggested that these two protein degradation inhibitors may be applicable for improving insect resistance in cotton production.

Marker-assisted selection to pyramid Fusarium head blight resistance loci Fhb1 and Fhb2 in the high-quality soft wheat cultivar Yangmai 15

Fusarium head blight(FHB)is one of the most detrimental wheat diseases which greatly decreases the yield and grain quality,especially in the middle and lower reaches of the Yangtze River of China.Fhb1 and Fhb2 are two major resistance loci against Fusarium graminearum.Yangmai 15(YM15)is one of the most popular varieties in the middle and lower reaches of the Yangtze River,and it has good weak gluten characters but poor resistance to FHB.Here we used Fhb1 and Fhb2 to improve the FHB resistance of YM15 by a molecular marker-assisted selection(MAS)backcrossing strategy.The selection of agronomic traits was performed for each generation.We successfully selected seven introgressed lines which carry homozygous Fhb1 and Fhb2 with significantly higher FHB resistance than the recurrent parent YM15.Three of the introgressed lines had agronomic and quality characters that were similar to YM15.This study demonstrates that the pyramiding of Fhb1 and Fhb2 could significantly improve the FHB resistance in wheat using the MAS approach.

Identification and fine mapping of a fertility restorer gene for wild abortive cytoplasmic male sterility in the elite indica rice non-restorer line 9311

The wild abortive(WA)-type cytoplasmic male sterility(CMS)derived from the wild rice species Oryza rufipogon Griff.is used widely in three-line indica hybrids.The identification and mapping of restorer of fertility(Rf)genes aided in the development of WA-type hybrids.Here we report that testcross F1 plants from the WA-type CMS line and 9311 exhibited stainable pollen grains with no seed set,indicating that 9311 carries minor-effect Rfs for WA-type CMS.We developed an advanced backcross population consisting of plants harboring small regions of donor chromosomal segments from 9311 in the WATianfeng A genetic background with moderate seed setting rates.Genetic analysis showed that the pollen fertility levels of the backcross individuals are governed by a single gene from 9311 that we named Rf19(t).By use of the RICE 40 K gene chip,three introduced segments were identified in the fertile lines,and a candidate region spanning 4.37–8.29 Mb on chromosome 1 was identified for Rf19(t).Finally,Rf19(t)was fine-mapped to a region of 90 kb between the DNA marker loci STS1-163 and STS1-183,in which eight ORFs were predicted.Also,using relative expression analyses,comparative sequence analyses and functional domain analyses,we identified LOC_Os01g10530 as the most likely candidate gene for Rf19(t).Furthermore,Rf19(t)was found to function in fertility restoration,most probably by regulating the degradation of m RNA transcribed from the mitochondrial gene WA352.These results increase our knowledge of fertility restoration in WA-type CMS lines and will facilitate the development of high-quality pairs of WAtype CMS and maintainer lines.

Decreased panicle N application alleviates the negative effects of shading on rice grain yield and grain quality

Light deficiency is a growing abiotic stress in rice production.However,few studies focus on shading effects on grain yield and quality of rice in East China.It is also essential to investigate proper nitrogen(N)application strategies that can effectively alleviate the negative impacts of light deficiency on grain yield and quality in rice.A two-year field experiment was conducted to explore the effects of shading(non-shading and shading from heading to maturity)and panicle N application(NDP,decreased panicle N rate;NMP,medium panicle N rate;NIP,increased panicle N rate)treatments on rice yield-and quality-related characteristics.Compared with non-shading,shading resulted in a 9.5-14.8%yield loss(P<0.05),mainly due to lower filled-grain percentage and grain weight.NMP and NIP had higher(P<0.05)grain yield than NDP under non-shading,and no significant difference was observed in rice grain yield among NDP,NMP,and NIP under shading.Compared with NMP and NIP,NDP achieved less yield loss under shading because of the increased filled-grain percentage and grain weight.Shading reduced leaf photosynthetic rate after heading,as well as shoot biomass weight at maturity,shoot biomass accumulation from heading to maturity,and nonstructural carbohydrate(NSC)content in the stem at maturity(P<0.05).The harvest index and NSC remobilization reserve of NDP were increased under shading.Shading decreased(P<0.05)percentages of brown rice,milled rice,head rice,and amylose content while increasing(P<0.05)chalky rice percentage,chalky area,chalky degree,and grain protein.NMP demonstrated a better milling quality under non-shading,while NDP demonstrated under shading.NDP exhibited both lower chalky rice percentage,chalky area,and chalky degree under non-shading and shading,compared with NMP and NIP.NDP under shading decreased amylose content and breakdown but increased grain protein content and setback,contributing to similar overall palatability to non-shading.Our results suggested severe grain yield and quality penalty of rice when subjected to shading after heading.NDP improved NSC remobilization,harvest index,and sink-filling efficiency and alleviated yield loss under shading.Besides,NDP would maintain rice’s milling,appearance,and cooking and eating qualities under shading.Proper N management with a decreased panicle N rate could be adopted to mitigate the negative effects of shading on rice grain yield and quality.

Salt Stress Affects the Growth and Yield of Wheat (Triticum aestivum L.) by Altering the Antioxidant Machinery and Expression of Hormones and Stress- Specific Genes

Understanding physiological responses in saline agriculture may facilitate wheat breeding programs.Based on a screening test,the Ningmai-14(NM-14)and Yangmai-23(YM-23)wheat cultivars were selected for further experiments to understand the underlying salinity tolerance mechanism.This study investigated the effects of five salinity levels such as Control(CK)=0(without NaCl stress),S1=0.20%,S2=0.25%,S3=0.30%and S4=0.35%of NaCl concentrations of soil on wheat plants.The results showed that increased salinity concentration reduced the growth and yield of wheat cultivars(NM-14 and YM-23).However,YM-23(12.7%)yielded more than NM-14 at maximum salinity stress.The higher salinity(S4)increased the concentration of Na^(+)(4.3 to 5.8-fold)and P contents(2.5 to 2.2-fold),while reducing the average concentrations of K^(+),Cu,and K^(+)/Na^(+)ratio.The higher salinity(S4)reduced the spikelet length by 21.35%(followed by grain spike−1),and the starch content by 18.81%.In the YM-23 cultivar,higher salinity increased superoxide dismutase(SOD),total antioxidant capacity(TAC),and amylase.Compared to NM-14,induced expression of TaYUC2,6,and TaGA13ox,20ox genes were recorded in YM-23.Similarly,in YM-23 the stress-specific genes such as TaHSP70,90 were enhanced whereas,TaSOS1,2 were suppressed.Overall,our study revealed that salt tolerant cultivars modulate hormonal and antioxidant activities,thus maintaining high growth.

The Effect of Organ Temperature on Total Yield of Transplanted and Direct-Seeded Rice(Oryza sativa L.)

The canopy temperature of rice is an important index that directly reflects the growth and physiological state of rice,and affects the yield of rice plants to a great extent.The correlation between the temperatures of different rice organs and canopy in different growth stages and the grain yield is complex.The stability and universality of these correlations must be verified.We conducted a pot experiment using two rice varieties and two temperature treatments(high temperature treatment was carried out at the beginning of heading stage for 10 days).We measured rice organ temperature during seven stages of growth using a high-precision infrared thermal imager.Results showed that the optimal observation period for the rice canopy temperature was 13:00.Although the rice variety did not significantly impact the canopy or organ temperature(p>0.05),the different organs and canopy exhibited significantly different temperatures(p<0.05).The correlations between the leaf,stem,panicle,canopy–air temperature differences and seed setting rate,theoretical and actual yields were the strongest during the milk stage.Among them,the correlation coefficient betweenΔT_(s) and theoretical and actual yields was the highest,the relationship between theoretical yield(Y)andΔT_(s)(X)was Y=−5.6965X+27.778,R^(2)=0.9155.Compared withΔT_(l),ΔT_(p) andΔTc,ΔT_(s) was closely related to the main traits of plants.ΔT_(s) could better reflect the growth characteristics of rice thanΔT_(c),such as dry matter accumulation(r=−0.931),SPAD(r=0.699),N concentration(r=0.714),transpiration rate(r=−0.722).In conclusion,stem temperature was more important indicator than canopy temperature.Stem temperature is a better screening index for rice breeding and cultivation management in the future.

Irrigation regimes modulate non-structural carbohydrate remobilization and improve grain filling in rice(Oryza sativa L.)by regulating starch metabolism

Recently developed ‘super’ rice cultivars with greater yield potentials often suffer from the problem of poor grain filling, especially in inferior spikelets. Here, we studied the activities of enzymes related to starch metabolism in rice stems and grains, and the microstructures related to carbohydrate accumulation and transportation to investigate the effects of different water regimes on grain filling. Two ‘super’ rice cultivars were grown under two irrigation regimes of well-watered(WW) and alternate wetting and moderate soil drying(AWMD). Compared with the WW treatment,the activities of ADP glucose pyrophosphorylase(AGPase), starch synthase(StSase) and starch branching enzyme(SBE), and the accumulation of non-structural carbohydrates(NSCs) in the stems before heading were significantly improved, and more starch granules were stored in the stems in the AWMD treatment. After heading, the activities of α-amylase, β-amylase, sucrose phosphate synthase(SPS) and sucrose synthase in the synthetic direction(SSs)were increased in the stems to promote the remobilization of NSCs for grain filling under AWMD. During grain filling, the enzymatic activities of sucrose synthase in the cleavage direction(SSc), AGPase, StSase and SBE in the inferior spikelets were increased, which promoted grain filling, especially for the inferior spikelets under AWMD.However, there were no significant differences in vascular microstructures. The grain yield and grain weight could be improved by 13.1 and 7.5%, respectively, by optimizing of the irrigation regime. We concluded that the low activities of key enzymes in carbon metabolism is the key limitation for the poor grain filling, as opposed to the vascular microstructures, and AWMD can increase the amount of NSC accumulation in the stems before heading, improve the utilization rate of NSCs after heading, and increase the grain filling, especially in the inferior spikelets, by altering the activities of key enzymes in carbon metabolism.