Genome-wide and candidate gene association studies identify BnPAP17 as conferring the utilization of organic phosphorus in oilseed rape

Phosphorus(P)is essential for living plants,and P deficiency is one of the key factors limiting the yield in rapeseed production worldwide.As the most important organ for plants,root morphology traits(RMTs)play a key role in P absorption.To investigate the genetic variability of RMT under low P availability,we dissected the genetic structure of RMTs by genome-wide association studies(GWAS),linkage mapping and candidate gene association studies(CGAS).A total of 52 suggestive loci were associated with RMTs under P stress conditions in 405 oilseed rape accessions.The purple acid phosphatase gene BnPAP17 was found to control the lateral root number(LRN)and root dry weight(RDW)under low P stress.The expression of BnPAP17 was increased in shoot tissue in P-efficient cultivars compared to root tissue and P-inefficient cultivars in response to low P stress.Moreover,the haplotype of BnPAP17^(Hap3)was detected for the selective breeding of P efficiency in oilseed rape.Over-expression of the BnPAP17^(Hap3)could promote the shoot and root growth with enhanced tolerance to low P stress and organic phosphorus(Po)utilization in oilseed rape.Collectively,these findings increase our understanding of the mechanisms underlying BnPAP17-mediated low P stress tolerance in oilseed rape.

Biotechnology ofα-linolenic acid in oilseed rape(Brassica napus)using FAD2 and FAD3 from chia(Salvia hispanica)

α-Linolenic acid(ALA,18:3Δ9,12,15)is an essential fatty acid for humans since it is the precursor for the biosynthesis of omega-3 long-chain polyunsaturated fatty acids(LC-PUFA).Modern people generally suffer from deficiency of ALA because most staple food oils are low or lack ALA content.Biotechnological enrichment of ALA in staple oil crops is a promising strategy.Chia(Salvia hispanica)has the highest ALA content in its seed oil among known oil crops.In this study,the FAD2 and FAD3 genes from chia were engineered into a staple oil crop,oilseed rape(Brassica napus),via Agrobaterium tumefaciens-mediated transformation of their LP4-2A fusion gene construct driven by the seed-specific promoter P_(NapA).In seeds of T0,T1,and T2 lines,the average ALA contents were 20.86,23.54,and 24.92%,respectively,which were 2.21,2.68,and 3.03 folds of the non-transformed controls(9.42,8.78,and 8.22%),respectively.The highest seed ALA levels of T0,T1,and T2 plants were 38.41,35.98,and 39.19%respectively,which were 4.10-4.77 folds of the respective controls.FA-pathway enzyme genes(BnACCD,BnFATA,BnSAD,BnSCD,BnDGAT1,BnDGAT2,and BnDGAT3)and positive regulatory genes(BnWRI1,BnLEC1,BnL1L,BnLEC2,BnABI3,BnbZIP67,and BnMYB96)were all significantly up-regulated.In contrast,BnTT1,BnTT2,BnTT8,BnTT16,BnTTG1,and BnTTG2,encoding negative oil accumulation regulators but positive secondary metabolism regulators,were all significantly down-regulated.This means the foreign ShFAD2-ShFAD3 fusion gene,directly and indirectly,remodeled both positive and negative loci of the whole FA-related network in transgenic B.napus seeds.

Ridge-furrow rainwater harvesting with supplemental irrigation to improve seed yield and water use efficiency of winter oilseed rape(Brassica napus L.)

Ridge-furrow rainwater harvesting(RFRH) planting pattern can lessen the effect of water deficits throughout all crop growth stages, but water shortage would remain unavoidable during some stages of crop growth in arid and semiarid areas. Supplemental irrigation would still be needed to achieve a higher production. Field experiments were conducted for two growing seasons(2012–2013 and 2013–2014) to determine an appropriate amount of supplemental irrigation to be applied to winter oilseed rape at the stem-elongation stage with RFRH planting pattern. Four treatments, including supplemental irrigation amount of 0(I1), 60 mm(I2) and 120 mm(I3) with RFRH planting pattern and a control(CK) irrigated with 120 mm with flat planting pattern, were set up to evaluate the effects of supplemental irrigation on aboveground dry matter(ADM), nitrogen nutrition index(NNI), radiation use efficiency(RUE), water use efficiency(WUE), and seed yield and oil content of the oilseed rape. Results showed that supplemental irrigation improved NNI, RUE, seed yield and oil content, and WUE. However, the NNI, RUE, seed yield and oil content, and WUE did not increase significantly or even showed a downward trend with excessive irrigation. Seed yield was the highest in I3 for both growing seasons. Seed yield and WUE in I3 averaged 3 235 kg ha^(–1) and 8.85 kg ha^(–1) mm^(–1), respectively. The highest WUE was occurred in I2 for both growing seasons. Seed yield and WUE in I2 averaged 3 089 kg ha^(–1) and 9.63 kg ha^(–1) mm^(–1), respectively. Compared to I3, I2 used 60 mm less irrigation amount, had an 8.9% higher WUE, but only 4.5 and 0.4% lower seed yield and oil content, respectively. I2 saved water without substantially sacrificing yield or oil content, so it is recommended as an appropriate cultivation and irrigation schedule for winter oilseed rape at the stem-elongation stage.

Assessment of genetic effects on important breeding traits in reciprocal DH populations of winter oilseed rape (Brassica napus L.)

Here we present a study on the genetic effects of agronomic and seed quality traits in double haploid(DH) populations,which were developed by microspore culture from reciprocal F1 hybrids produced between cultivar Californium and DH W-15.Each of the populations consisted of 25 DH lines.The field experiments were conducted in a randomized complete block design with three replications during three growing seasons.Field observations were recorded on seed yield and its structure: the number of branches and pods per plant,the number of seeds per pod,the 1 000-seed weight,the content of fat in the seeds,and three unsaturated fatty acids(oleic,linoleic,linolenic) in the seed oil.In order to investigate the influence of cross direction on the studied traits,parental effects were evaluated on the basis of differences between reciprocal DH populations.The maternal effect was revealed on the number of seeds per pod and the effect of the paternal form on linolenic acid content.The occurrence of transgression effects also depended on the direction of crossing and this was particularly noticeable in terms of the number of seeds per pod.The use of multivariate statistical methods allowed for the simultaneous characterization and grouping of tested lines in terms of several traits.Graphic images of the distribution of DH lines in the space of the two first canonical variates showed a great variation between the two reciprocal populations,both in terms of yield and its components,as well as fat and those unsaturated fatty acids.

Utilizing comprehensive decision analysis methods to determine an optimal planting pattern and nitrogen application for winter oilseed rape

Oilseed rape is one of the most important oil crops globally.Attaining the appropriate cultivation method(planting pattern and nitrogen level)is necessary to achieve high yield,quality and resource utilization efficiency.However,the optimal method for oilseed rape varies across countries and regions.The objective of the present study was to determine an appropriate cultivation method,including planting pattern and nitrogen application,for winter oilseed rape in northwestern China.Two planting patterns:ridge film mulching and furrow planting(RFMF)and flat planting(FP),and six nitrogen(N)amounts:0(N0),60(N60),120(N120),180(N180),240(N240),and 300(N300)kg N ha–1 were applied across three growing seasons(2014–2017).Three comprehensive decision analysis methods:principal component analysis,grey correlation degree analysis and the combined entropy weight and dynamic technique for order preference by similarity to ideal solution method were used to evaluate the growth and physiological indicators,nutrient uptake,yield,quality,evapotranspiration,and water use efficiency of winter oilseed rape.Planting pattern,nitrogen amount and their interaction significantly affected the indicators aforementioned.The RFMF pattern significantly increased all indicators over the FP pattern.Application of N also markedly increased all the indicators except for seed oil content,but the yield,oil production and water use efficiency were decreased when N fertilizer exceeded 180 kg N ha–1 under FP and 240 kg N ha–1 under RFFM.The evaluation results of the three comprehensive decision analysis methods indicated that RFMF planting pattern with 240 kg N ha–1 is an appropriate cultivation method for winter oilseed rape in northwestern China.These findings are of vital significance to maximize yield,optimize quality and improve resource use efficiencies of winter oilseed rape.

Study of Spectral Response Characteristics of Oilseed Rape (Brassica napus) to Particulate Matters Based on Hyper-Spectral Technique

Haze is mainly caused by the suspended particulate matters in the air,of which the particulate matters pollution harms leaf vegetables.In this paper,oilseed rapes at four different growing periods were investigated in a simulated particulate pollution environment.In combination of hyper-spectral technology and micro examination,the response of hyper-spectral characteristics of the leaf to particulate matters was investigated in-depth.The hyperspectral,chlorophyll content,net photosynthetic rate and stomatal conductance of leaf were obtained.The deposition and adsorption of particulate matters on the leaf were observed by Environmental Scanning Electron Microscope(ESEM).Normalized difference vegetation index(NDVI),modified red edge normalized(mNDVI705)and modified red edge simple ratio index(mSR705)were selected as characteristic parameters and the range of 510 nm~620 nm as the sensitive band.16 methods were used to establish the physiological information inversion model.The main results were as follows:Under the influence of particulate matters,the spectral reflectance decreased as a whole.With the increase of leaf age,the phenomenon of blue shift aggravated.The amplitude of yellow and blue edge decreased with overall decreasing vegetation indices.The furrows and irregular band protrusions in leaves were favorable for keeping particulate matters.With longer affecting time and more deposition of particle matters on the leaf,the stomatal opening became smaller.After comparing,principal component regression(PCR)+multiple scatter correction(MSC)+second derivative(SD)+Savitzky-Golay smooth(SG),and partial least square(PLS)+multiple scatter correction(MSC)+first derivative(FD)+Savitzky-Golay smooth(SG)were determined the best method to establish the inversion model of chlorophyll content and net photosynthetic rate respectively.This study may bring novel ideas for the diagnosis and analysis of the physiological response of leaf vegetables under particulate matters pollution using hyper-spectral technology.

Co-exposure of sulfur nanoparticles and Cu alleviate Cu stress and toxicity to oilseed rape Brassica napus L.

Experimentswere performed to explore the impact of sulfur nanoparticles(SNPs)on growth,Cu accumulation,and physiological and biochemical responses of oilseed rape(Brassica napus L.)inoculated with 5 mg/L Cu-amended MS medium supplemented with or without 300 mg/L SNPs exposure.Cu exerted severe phytotoxicity and inhibited plant growth.SNPs application enhanced the shoot height,root length,and dry weight of shoot and root by 34.6%,282%,41.7%and 37.1%,respectively,over Cu treatment alone,while the shoot and root Cu contents and Cu-induced lipid perodixation as the malondialdehyde(MDA)levels in shoots and roots were decreased by 37.6%,35%,28.4%and 26.8%.Further,the increases in superoxide dismutase(SOD),peroxidase(POD),catalase(CAT),ascorbate peroxidase(APX),glutathione reductase(GR)and glutathione S-transferase(GST)enzyme activities caused by Cu stress were mitigated in shoots(10.9%–37.1%)and roots(14.6%–35.3%)with SNPs addition.SNPs also positively counteracted the negative effects on shoot K,Ca,P,Mg,Mn,Zn and Fe contents and root K,Ca,Mg and Mn contents from Cu exposure alone,and significantly promoted the nutrients accumulation in plant.Additionally,in comparison with common bulk sulfur particles(BSPs)and sulfate,SNPs showed more positive effects on promoting growth in shoots(6.7%and 19.5%)and roots(10.9%and 15.1%),as well as lowering the shoot Cu content(40.1%and 43.3%)under Cu stress.Thus,SNPs application has potential to be a green and sustainable technology for increasing plant productivity and reducing accumulation of toxic metals in heavy metal polluted soils.

Host-induced silencing of MpPar6 confers Myzus persicae resistance in transgenic rape plants

Plant-mediated RNA interference(RNAi)has emerged as a promising technology for insect control.The green peach aphid,Myzus persicae,feeds on over 400 species of host plants.Brassica napus(rape)is the second most important oilseed crop worldwide.Myzus persicae is highly reproductive and causes severe damage to the rape plants due to its quite flexible life cycle.In this study,we tested the RNAi effects of transgenic rape plants on M.persicae.By in vitro feeding M.persicae with artificial diets containing double-stranded RNAs(dsRNAs)targeting seven aphid genes,we identified a new gene encoding the partitioning-defective protein 6(Par6)as the most potent RNAi target.Tissue-and stage-expression analysis of Par6 suggested this gene is highly expressed in the embryo and adult stage of M.persicae.We next generated transgenic rape plants expressing ds Par6 by Agrobacteriummediated transformation and obtained nine independent transgenic lines.Compared to wild-type control plants,transgenic rape lines expressing ds Par6 showed strong resistance to M.persicae.Feeding assays revealed that feeding transgenic rape plants to M.persicae significantly decreased MpPar6 expression and survival rate and impaired fecundity.Furthermore,we showed that the resistance levels to M.persicae are positively correlated with ds Par6 expression levels in transgenic rape plants.Our study demonstrates that transgenic rape plants expressing ds Par6 are efficiently protected from M.persicae.Interfering with the genes involved in embryo development could be the effective RNAi targets for controlling aphids and potentially other insect pests.

Assessing and characterizing oilseed rape freezing injury based on MODIS and MERIS data

The oilseed rape growing in the lower reaches of Yangtze River in China belongs to winter varieties and suffers the risk of freezing injury.In this research,a typical freezing injury event occurred in Anhui Province was taken as a case study,the freezing damage degree of oilseed rape was assessed,and its development characteristics based on the vegetation metrics derived from MODIS and MERIS data were investigated.The oilseed rape was mapped according to the decline of greenness from bud stage to full-bloom period,with the phenological phases identified adopting time-series analyses.NDVI was more sensitive to freezing injury compared with other commonly used vegetation indices(VIs)calculated using MODIS bands,e.g.,EVI,GNDVI and SAVI.The freezing damage degree employing the difference between post-freeze growth and the baseline level in adjacent damage-free growing seasons was determined.The remote sensing-derived damage levels were supported by their correlation with the cold accumulated temperatures at the county level.The performance of several remote sensing indicators of plant biophysical and biochemical parameters was also investigated,i.e.,the photosynthetic rate,canopy water status,canopy chlorophyll content,leaf area index(LAI)and the red edge position(REP),in response to the advance of the freezing damage.It was found that the photosynthetic rate indicator—Photochemical Reflectance Index(PRI)responded strongly to freezing stress.Freezing injury caused canopy water loss,which could be detected though the magnitude was not very large.MERIS-LAI showed a slow and lagging response to low temperature and restored rapidly in the recovery phase;additionally,REP and the indicator of canopy chlorophyll content—MERIS Terrestrial Chlorophyll Index(MTCI),did not appear to be influenced by freezing injury.It was concluded that the physiological functions,canopy structure,and organic content metrics showed a descending order of vulnerabilities to freezing injury.

De novo design of future rapeseed crops:Challenges and opportunities

To address the global demand for rapeseed while considering farmers’profit,we face the challenges of making a quantum leap in seed yield and,at the same time,reducing yield loss due to biotic and abiotic stresses.We also face the challenge of efficiently applying new transformative biotechnology tools such as gene editing and breeding by genome design to increase rapeseed productivity and profitability.In this Perspective,we review advances in research on the physiological and genetic bases of both stress factorsaffected yield stability and seed yield potential,focusing on source–sink relationships and allocation of photosynthetic assimilates to vegetative growth and seed development.We propose research directions and highlight the role of plant architecture in the relative contributions of the root system,leaves,and pods to seed yield.We call for de novo design of new rapeseed crops.We review trait variation in existing germplasm and biotechnologies available for crop design.Finally,we discuss opportunities to apply fundamental knowledge and key germplasm to rapeseed production and propose an ideotype for de novo design of future rapeseed cultivars.

Effects of different rotation patterns on the occurrence of clubroot disease and diversity of rhizosphere microbes

Clubroot disease, caused by Plasmodiophora brassicae, is one of the most destructive soil-borne diseases in cruciferous crops worldwide. New strategies are urgently needed to control this disease, as no effective disease-resistant varieties or chemical control agents exist. Previously, we found that the incidence rate and disease index of clubroot in oilseed rape decreased by 50 and 40%, respectively, when oilseed rape was planted after soybean. In order to understand how different rotation patterns affect the occurrence of clubroot in oilseed rape, high-throughput sequencing was used to analyze the rhizosphere microbial community of oilseed rape planted after leguminous (soybean, clover), gramineous (rice, maize) and cruciferous (oilseed rape, Chinese cabbage) crops. Results showed that planting soybeans before oilseed rape significantly increased the population density of microbes that could inhibit P. brassicae (e.g., Sphingomonas, Bacillus, Streptomyces and Trichoderma). Conversely, consecutive cultivation of cruciferous crops significantly accumulated plant pathogens, including P. brassicae, Olpidium and Colletotrichum (P<0.05). These results will help to develop the most effective rotation pattern for reducing clubroot damage.

Molecular regulation and genetic improvement of seed oil content in Brassica napus L.

As an important oil crop and a potential bioenergy crop, Brassica napus L. is becoming a model plant for basic research on seed lipid biosynthesis as well as seed oil content, which has always been the key breeding objective. In this review, we present current progress in understanding of the regulation of oil content in B. napus, including genetics, biosynthesis pathway,transcriptional regulation, maternal effects and QTL analysis. Furthermore, the history of breeding for high oil content in B. napus is summarized and the progress in breeding ultra-high oil content lines is described. Finally,prospects for breeding high oil content B. napus cultivars are outlined.

Stability evaluation of the PROSPECT model for leaf chlorophyll content retrieval

The radiative transfer model,PROSPECT,has been widely applied for retrieving leaf biochemical traits.However,little work has been conducted to evaluate the stability of the PROSPECT model with consideration of multiple factors(i.e.,spectral resolution,signal-to-noise ratio,plant growth stages,and treatments).This study aims to investigate the stability of the PROSPECT model for retrieving leaf chlorophyll(Chl)content(Cab).Leaf hemispherical reflectance and transmittance of oilseed rape were acquired at different spectral resolutions,noise levels,growth stages,and nitrogen treatments.The Chl content was also measured destructively by using a microplate spectrophotometer.The performance of the PROSPECT model was compared with a commonly used random forest(RF)model.The results showed that the prediction accuracy of PROSPECT and RF models for Cab did not produce significant differences under varied spectral resolutions ranging from 1 to 20 nm.The ranges of the relative root mean square errors(rRMSE)of the PROSPECT and RF models were 12%-13%and 11.70%-12.86%,respectively.However,the performance of both models for leaf Chl retrieval was strongly influenced by the noise level with the rRMSE of 13-15.37%and 12.04%-15.80%for PROSPECT and RF,respectively.For different growth stages,the PROSPECT model had similar prediction accuracies(rRMSE=9.26%-12.41%)to the RF model(rRMSE=9.17%-12.70%).Furthermore,the superiority of the PROSPECT model(rRMSE=10.10%-12.82%)over the RF model(rRMSE=11.81%-15.47%)was prominently observed when tested with plants growth at different nitrogen treatment levels.The results demonstrated that the PROSPECT model has a more stable performance than the RF model for all datasets in this study.