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.

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.

Strategies and Economic Benefit Analysis for Productive Landscape of Rape Flower Sea in Cold Regions

The cold regions are located in high latitudes and cold climates.The local excellent ornamental plant resources are relatively scarce.The plant species that are suitable for both ornamental and productive benefits of landscape of flower sea construction are also even fewer.Therefore,it is imperative to introduce and screen the plant resources suitable for cold regions to create the landscape of flower sea.The rape,an oilseed crop,was used as a research object in order to create a productive flower landscape with both ornamental and economic values in cold regions.Four rape flower varieties,Qingza No.5,7,9,and 11,were introduced from Qinghai Hufeng Agricultural Science and Technology Group Co.,Ltd.They were planted in the experimental practice base of Northeast Agricultural University in three batches.Development characteristics and seed yield of rape flowers on different sowing dates were studied.The fuzzy probability method was used to comprehensively evaluate the varieties.The results showed that the rape flowers grew well in Harbin City during the experimental sowing period,which could form a good landscape of flower sea and had a considerable rapeseed yield.It could be widely used in cold urban and rural areas,such as Harbin City.In view of the experimental results,the strategies of creating a productive landscape of rape flower sea were proposed and the economic benefits were analyzed.It could change the status quo of a uniform landscape of flower sea in cold regions,help the development of rural tourism,and promote local economic income.

Different Foliar Fertilizer Treatments Affect the Yield and Quality of Brassica napus L.for Both Vegetable and Oil Dual-Use Rape

Field experiments were carried out to study the effect of different foliar fertilizer treatments on the agronomic traits,yield,and quality of Brassica napus L.for both vegetable and oil dual-use rape.The seedlings of‘Youtai 929’,‘Jinyoutai 1’,and‘Zhongshuang 11’were sprayed with potassium dihydrogen phosphate,naphthylacetic acid,and selenium fertilizer,and the agronomic traits,yield,and quality were determined.The results showed that the naphthylacetic acid treatment had the highest indexes before winter,followed by the selenium fertilizer treatment.At the harvesting stage,the plant height,number of siliquae,yield per plant,and number of effective branches on the main stem of the three varieties were the highest in the naphthylacetic acid treatment,followed by those in the potassium dihydrogen phosphate treatment.The fresh shoot quality was the best in the selenium fertilizer treatment and the second in the naphthylacetic acid treatment.‘Jinyoutai 1’applied with naphthylacetic acid had the highest economic benefit of 4971.78 CNY/667m2,which was 642.32 CNY/667m2 higher than that of the control.The findings suggested that naphthylacetic acid was suitable to be used as the foliar fertilizer for the seedlings of B.napus varieties for both vegetable and oilseed,and the additional application of selenium fertilizer demonstrated better effect.

Rape Breeding and Its Intellectual Property Protection in China

Rape is the third largest oil crop in the world and the first one in China.The national average rapeseed yield is only 2077.5 kg/hm^(2),and the rapeseed oil yield is 894 kg/hm^(2).This paper introduced the main types,germplasm resources,breeding bases and breeding progress of Chinese rape,and studied its high-quality varieties,breeding patents and new varieties of agricultural plants.We analyzed the main problems such as serious loss of germplasm resources in Brassica campestris L.and Brassica juncea(L.)Czern.et Coss and weak"agricultural chip"in rape field,and put forward strategies such as strengthening the protection of rape germplasm resources,encouraging the development of"agricultural chip"in rape industry clusters with national advantages and characteristics,and strengthening the protection of agricultural intellectual property rights.

Geographical Indication Intellectual Property Protection and Regional Public Brand Construction of Rape Industry in China

As China s largest oil crop,rape occupies a central position in ensuring the safety of China s cooking oil supply.This paper introduced China s rapeseed industry from the rape type,rapeseed production,planting area,national rapeseed production protected area,rape national dominant characteristic industrial clusters,and rapeseed industry,etc.Besides,from the aspects of geographical indication products,geographical indication trademarks,and geographical indications of agricultural products,this paper discussed the intellectual property protection of geographical indications of rape,rapeseed,and rapeseed oil in China.It analyzed the main problems such as the lag in the formulation of relevant standards for geographical indications and the low use of special signs for geographical indications,and finally came up with recommendations including building a public brand of geographical indications and expanding foreign exchanges of geographical indications.

National Protection Zone for Rapeseed Production and Industrial Cluster of Rape in Hubei Province

Hubei Province is the main winter rapeseed producing area in China,and has the national advantage and characteristic industrial cluster of rape.The protection zone for the production of important agricultural products of rapeseed in Hubei Province also ranks first as that in Hunan Province.This paper studied the winter rape industry,the important agricultural product production protection zone of rapeseed,the provincial double-low high-quality rape protection zone and the industrial cluster of rape in Jianghan Plain in Hubei Province,and analyzed the main problems existing in the development of rapeseed production protection zone and industrial cluster of rape.Finally,it came up with strategies including promoting national rape varieties,overcoming the problem of rape"agricultural chip",promoting the"rice+rape"rotation model,and strengthening the protection of intellectual property rights of brand marks.

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.

An Artificial Approach for the Fractional Order Rape and Its Control Model

The current investigations provide the solutions of the nonlinear fractional order mathematical rape and its controlmodel using the strength of artificial neural networks(ANNs)along with the Levenberg-Marquardt backpropagation approach(LMBA),i.e.,artificial neural networks-Levenberg-Marquardt backpropagation approach(ANNs-LMBA).The fractional order investigations have been presented to find more realistic results of the mathematical form of the rape and its control model.The differential mathematical form of the nonlinear fractional order mathematical rape and its control model has six classes:susceptible native girls,infected immature girls,susceptible knowledgeable girls,infected knowledgeable girls,susceptible rapist population and infective rapist population.The rape and its control differential system using three different fractional order values is authenticated to perform the correctness of ANNs-LMBA.The data is used to present the rape and its control differential system is designated as 70%for training,14%for authorization and 16%for testing.The obtained performances of the ANNs-LMBA are compared with the dataset of the Adams-Bashforth-Moulton scheme.To substantiate the consistency,aptitude,validity,exactness,and capability of the LMBA neural networks,the obtained numerical values are provided using the state transitions(STs),correlation,regression,mean square error(MSE)and error histograms(EHs).

Maternal-Fetal Pregnancy Complications of in Minors Raped with or without Physical Restraint in Eastern Democratic Republic of Congo

Introduction: Pregnancy resulting from rape is a public health and sexual and reproductive health issue, especially among minors. Rape can be perpetrated with or without physical restraint. The objective of the present study is to highlight the prevalence of physical coercion leading to pregnancy and the associated maternal-fetal complications. Methodology: This was a comparative descriptive and analytical cross-sectional study of complications associated with post-rape pregnancy by physical restraint among minors who were treated at the General Reference Hospital of Panzi over a two-year period from June 2020 to June 2022. A total of 140 minor survivors of violence with pregnancy were included in the study. Data were collected using a questionnaire, coded and analyzed in Excel and XLSTAT version 2014. Results: The prevalence of physical constraints was 65%. After mulltivariate analyses, denial of pregnancy (ORa: 9.64 95% CI: 1.1 - 81.2;p-value: 0.0370), attempted abortion (ORa: 56.1 95% CI: 1.5 - 2027.6;p-value: 0.0278) and agitation during delivery (ORa: 88.7 95% CI: 4.5 - 1715;p-value: 0.0030) were the complications associated with pregnancy in minors who experienced physical restraint rape. In addition, BMI was a factor in reducing the risk of physical restraint rape at the ORa of 0.5054 [0.3;0.8];p 0.006). Conclusion: Pregnancy among minors is a reality and occurs in a situation of physical coercion in the eastern Democratic Republic of Congo. These pregnancies are often associated with complications that require an intensive management system since they can jeopardize the maternal-fetal prognosis of minors.

Analysis of Correlation between Yield and Quality Traits of Quality Hybrid Rapeseed

[Objective] This study aimed to investigate the correlation between yield and quality traits of rape. [Method] Taking ＂Sanbei 98＂ and ＂Youyan 599＂ as ex- periment materials, correlation between yield and quality traits of Brassica napus L. cultivars was explored by orthogonal rotation method. [Result] The results showed that： 1 In both the two varieties-Sanbei98 and Youyan599, erucic acid content had the largest coefficient of variation, but its average content was lower than 2% in both the two varieties （0.90% in Sanbei98 and 1.24% in Youyan599）, and the palmitic acid content had the smallest coefficient of variation. 2 In terms of the dif- ferences in quality traits at different yield levels, only the mean of eicosenoic acid content revealed significant difference in Sanbei98, and only the mean of seed-pro- tein content revealed significant difference in Youyan599, and there was no signifi- cant difference in any combined quality traits in both the two varieties. 3 The yield was negatively correlated with the erucic acid content or oil content, and posi- tively correlated with the content of seed-protein, linoleic acid, or linolenic acid, and the coefficient of correlation between the yield and either of the contents of glucosi- nolates, oleic acid, palmitic acid, stearic acid, and eicosenoic acid was smaller. ~, when the yield of a variety increased considerably, there was a certain decreasing trend in erucic acid content, oil content, the total amount of erucic chain fatty acids （El ＆ Er）, while there was an increasing trend in seed-protein content, linoleic acid content, linolenic acid content, saturated fatty acids （SFA）, the total amount of fat and protein （P ＆ F）, unsaturation index （IUFA）, unsaturated fatty acids （UFA）, the total amount of 18 carbon fatty acids （18-C FA）, unsaturated fatty acids [UFA/（Ei ＆ Er） ], and the sum of oleic acid and linoleic acid （OI ＆ Li）. 5 When the seed-pro- tein content is increased, the quality of rape oil cake is also improved; when the unsaturated index is increased, the oil stability is decreased. The increase in the total amount of unsaturated fatty acids, oleic acid and linoleic acid helps to increase in the total amount of fatty acids absorbed by human bodies. [Conclusion] This study provides a theoretical foundation for the cultivation of high-yielding and top- quality hybrid rape cultivars.

Research on the Prevention and Control Technology of Rape Aphids

[Objective] The aim was to screen economic and environment-friendly pesticides adaptive to aphid-resistant varieties in main rape growing regions of Yunnan Province.[Method] The effects of 6 pesticides against aphids on 22 Brassica campestris materials of the main breeds in Yunnan Province,the new improved varieties in China and the core breeding materials were tested.[Result] Yunyoushuang 1,Huayou 4,Yunhuayou Early-maturing Variety No.1 and A35 showed better aphid-resistance.An optimum pesticide application strategy was to use Diyaling in sowing time,and interchangeably apply nitenpyram and imidacloprid during flowering and pod formation stages.[Conclusion] The present study had provided an important technical support for sustainable development of rape industry.

Chloroplast Composition and Structural Differences in a Chlorophyll-reduced Mutant of Oilseed Rape Seedlings

对黄化突变体Cr352 9和野生型油菜 (BrassicanapusL .) 352 9叶绿体的超微结构和组成进行了比较。与野生型相比 ,突变体Cr352 9叶片具有较少的类囊体、较少的垛叠膜区和较少的叶绿素含量。突变体的Chla/Chlb比值较高 ,是野生型的 2倍。电泳结果表明 ,突变体类囊体膜中LHCⅡ和其三聚体LHCⅡ 的含量减少。SDS_PAGE分析显示 ,LHCⅡ的脱辅基蛋白在突变体类囊体膜中明显减少。免疫印迹进一步表明 ,所有LHCⅡ组分的含量仅为野生油菜的类囊体膜的 1 / 3。突变体Cr352 9的天线系统比野生型 352 9的小。

Effects of Nitrogen Application Rate, Density and Seedling Age on Dry Matter Accumulation of No-tillage Rape in Seedling Stage

[Objective] The aim of the research was to find the optimal nitrogen application rate, density and seedling age for no-tillage rape in seedling stage. [Method] With the D-optimal quadratic regression design for three factors, the 310 scheme was designed to study the effects of nitrogen application rate, density and seedling age on dry matter accumulation of no-tillage rape in seedling stage. [Result] With the increase of nitrogen application rate, density and seedling age, the dry matter content appeared like a parabola, increasing firstly and then declining. The change of nitrogen application rate caused greater influence than that of density and seedling age; the interaction effects between nitrogen application rate and density were greater than that between nitrogen application rate and seedling age as well as between density and seedling age. [Conclusion] Considered comprehensively, the dry matter content of no-tillage rape in seedling stage reached the highest level (4 768.2 kg/hm2) when the nitrogen application rate, the density and the seedling age were 195 kg/hm2, 93 000 plants/hm2 and 33 d, respectively.

Biomechanical Characteristics of Rape Stalks

[Objective] To study the correlation between the biomechanical properties of rape stalks and rape stem lodging. [Method] Through axial compression tests to the stalks of 4 different rape varieties, the change rules of maximum stem bearing ca- pacity, maximum compressive strength, elastic modulus and moment of inertia along plant height were analyzed, as well as the effect of different varieties and water contents on the biomechanical property indices of rape stalks. [Result] The maximum loads of rape stalks presented liner decrease trend along with the increase of stem height, and all reached the maximums below the height of 50 cm. The maximum stem compressive strength and elastic modulus of the 4 varieties were increased with ascending height, but in a slow rate with small change, thus the modulus of e- lasticity could be considered as unchanged. The maximum bearing capacity, maxi- mum compressive strength and elastic modulus of dry rape stalks were higher than wet stalks, indicating that the water contents of rape stalks had significant effect on their mechanical properties. According to the actual lodging situations in filed, stalks of variety No. 1 owned the worst biomechanical properties and lodging degree, while the biomechanical properties of No. 6 and F5 were better than No. 1 and No. 9, and they also had stronger lodging-resistance. [Conclusion] The study provides parameters and bases for the design of mechanized production and mechanical deep processing of crops, and can better reveal the physical natures of organisms. The methods used in this study can also be used to screen excellent crop stalks.

Dynamics of Essential Metal Elements Contents in Winter Oilseed Rape during Cold Acclimation

[Objective]The paper was to investigate the dynamics of essential metal elements（Ca,Mg,Fe,Mn,Zn and Cu）contents in winter oilseed rape（Brassica napus L.）during cold acclimation,and to reveal the effects and mechanisms of essential elements in cold resistance of oilseed rape.[Method]Three varieties with different cold resistance including Zhongshuang No.11（ZS,freezing sensitive variety）,Ganyouza No.1（GY）and Jinyuyou No.1（JYY,freezing tolerant varieties）were used in the study,and the changes of necessary metal elements in these oilseed rapes during cold acclimation were studied.[Result]The concentration of metal elements in oilseed rape had significant changes during cold acclimation,and the difference between varieties was significant.In plant roots,the concentrations of Ca,Mg,Fe,Mn,Zn and Cu were significantly increased,this might because that low temperature made the transpiration decreased,thus blocking the upward transport of elements.In plant shoots,the concentrations of Ca,Mg and Zn were significantly decreased,while the concentrations of Fe and Mn were significantly increased;the change extent of various elements in ZS variety was the largest.[Conclusion]Maintaining the stability of Ca,Mg and Zn concentrations and increasing Fe,Mn and Cu concentrations in plant shoots might be benefit for increasing the cold tolerance of winter oilseed rape.

Effect of Different Nitrogen Application Amount on the Net Increment of Fresh Dry Weight in Hybrid Rape

[ Objective] The aim of the research was to provide reference for reasonable application of nitrogen fertilizer for high yield.cultivation of hybrid rape cuhivar Youyan 9 and Youyan 10. [ Method] The net increment changes of individual plant fresh weight and dry matter weight of Youyan 9 and Youyan 10 with different nitrogen application treatments were studied. [ Result] The differences among average fresh weight increments of individual plant and average dry matter weight increment of individual plant with different treatments reached 0. 01 extremely significant level. Fresh weight increment and dry matter weight net increment of individual plant declined gradually with the increase of nitrogen application. In growtheourse ,fresh weight net increment of individual plant increased firstly then decreased and the maximum was in beginning flowering stage, besides that dry matter net increment increased gradually and the maximum was in mature period. The correlations among fresh net increment, dry matter weight net increment and yield net increment were positive or extremely positive. [ Conclusion] Under experimental condition, when nitrogen application was 225 kg/hm^2, hybrid rape Yanyou 9 and Yanyou 10 with low erucic,low glucosinolate could obtain high yield.

Research on the Production Efficiency and Utilizing Rate of Potassium Fertilizer for Rape in Yunnan Province

[Objective] This study aimed at investigating the effects of consumption of potassium fertilizer on production and biological characteristics of rape and utilizing efficiency of potassium fertilizer.[Method] Taking 0 kg/hm2 as control,nine consumptions of potassium fertilizer were designed for the experiment using single factor randomized block design with three replications and field experiment in nine rape-producing areas with different ecological types in Yunnan Province.[Result] Appropriately applying potassium fertilizer in Yunnan Province could improve the rapeseed production,the rape production could achieve 3 983-4 151 kg when applied 150-180 kg/hm2 of K2O,which had increased by 4.7-9.1% compared with the control and attained the peak in K165 treatment.When applied 1 kg of K2O,the rapeseed production could enhance 559 kg,the net profit could reach 1 229 yuan/hm2 and the partial productivity of potassium fertilizer could attain 31.32 kg/kg K2O and 3.7 kg/kg K2O,respectively.Applying potassium fertilizer could promote the growth of rape and increase rapeseed production.[Conclusion] The recommended optimal consumptions of potassium fertilizer with high production and high efficiency for rape in Yunnan Province were ranged between 150 and 180 kg K2O/hm2.However,comprehensively considering production and efficiency factors,the consumption of potassium fertilizer in Yunnan Province was around 73.9 kg/hm2,which should be appropriately increased to around 100.0 kg/hm2 in Longchuan,Yulong,Linxiang and other producing areas with similar conditions.

Effects of Sludge Application on Accumulition of Heavy Metals in Rape and Soil

[Objective] This study aimed to provide reliable theoretical basis for re- source utilization of sludge. [Method] By using pot experiment, variations of the yield index and salt content of rape and the accumulation of three kinds of heavy metals in rape plants and soil including Hg, As and Zn were explored. [Result] The results showed that medium sludge level （50 g/pot） significantly increased the rape yield and root/shoot ratio; when the application amount of sludge reached 100 g/pot, the fertil- ization effect of sludge on potted rape yield was significantly reduced. Compared with chemical fertilizer and chicken manure, sludge promoted N and P absorption of rape more significantly, but the effect of sludge application on K absorption was less significant than chicken manure. Sludge can increase the soil salinity, which is higher than the chemical fertilizer and chicken manure, but the effect was not significant. Compared with chemical fertilizer and chicken manure treatments, the Hg, As and Zn contents in rape plants and soil after sludge application increased, to be specific, Hg content increased relatively significantly. Hg, As and Zn contents in rape plants and soil were all in accordance with China National Safety Standards for Heavy Met- als in Vegetables and China National Environmental Quality Standards for Soils. [Conclusion] Short-term sludge application will not lead to secondary heavy metal pollution of agricultural soils, but the Hg content should be monitored and controlled in long-term sludge application.

Effects of Spraying KCl Solution on Photosynthate and Grain Yield and Quality of Oilseed Rape(Brassica napus L.)

[Objective] In order to seek the way of high quality and high yield of oilseed rape.[Method] Soil culture experiment was used in this research.1% （mass fraction） KCl solution was sprayed to the stems and leaves of oilseed rape in full blooming stage while 1.17% （mass fraction） K2SO4 solution and clear water were used for comparison,sampling in initial silique stage and harvest stage were conducted to detect photosynthate,soluble sugar and free amino acids content.[Result] The results showed that compared with spraying clear water,spraying KCl solution increased the contents of chlorophyll a in functional leaves,the contents of free amino acids and soluble sugar in leaves,stems and silique,and the total amounts of them all in initial silique stage but decreased the contents of free amino acids and soluble sugar in leaves and stems,and the total amounts of them all in harvesting stage,increased the average grain yield 10.20%,increased the average contents of oil,oleinic acid and linoleic acid in rapeseed 4.40%,10.60% and 11.40% respectively,decreased the content of protein,erucic acid and glucosinolate in rapeseed 5.10%,9.70% and 3.70% respectively.No significant difference between spraying K2SO4 solution and spraying clear water,significant difference between them all and spraying KCl solution.[Conclusion] The experimental results provided theoretical basis for increasing oilseed rape yield and quality researches.