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.

The Relationship of Color Formation with Related Enzymes and Protein Contents in the Seedcoat of Oilseed Rape (Brassica napus)

Three pairs of near-isogenic lines with different genetic backgrounds of yellow-seededand black-seeded rape (Brassica napus L.) were used as experiment materials to study therelationship of color formation in the seedcoat with enzyme activity and protein contentin it. The results showed that with similar genetic backgrounds, phenylalanine ammonia-lyase (PAL) and polyphenol oxidase(PPO) activities in the black-seeded lines were muchhigher than in their yellow-seeded counterparts and maximum PAL activity in the seedcoatoccurred comparatively late while no significant difference was present in glutaminesynthetase (GS) between the two types of rape. The plants were treated with red light,blue light, p-hydroxybenzoic acid (a PAL inhibitor), polyvinylpyridoxal (a PPO inhibitor),urea (a protein synthesis promoter) or chloramphenicol (CM, a plastid protein synthesisinhibitor) during seed development. It is speculated that PAL may be primarily responsiblefor coloration in the yellow seed; PPO may be the main factor contributing to thedarkness of the testa of the black genotypes; and nitrogen assimilation is, probably, notdirectly related to the difference in protein content observed between yellow- andblack-seeded genotypes, which may be induced mainly by PAL.

Mechanical Harvesting Effects on Seed Yield Loss,Quality Traits and Profitability of Winter Oilseed Rape (Brassica napus L.)

China is one of the most important rapeseed producing countries in the world. Effective mechanical harvesting time for decreasing harvesting loss of winter oilseed rape has been becoming a critical factor. An elite cultivar Zhongshuang 11 （Brassica napus L.） was employed in two rounds of field experiments from 2009 to 2011. Seeds were sown with machine, three combine harvesting times namely combine harvesting A, B, and C （CHA, CHB, and CHC） were designed and manual harvesting （MH） as control was performed at maturity. The harvesting treatments were determined according to color of pod and seed in the field. Seed yield loss and quality in different treatments were evaluated. Results showed that both seed yields and harvesting losses in 2009-2010 were higher than that in 2010-2011, whereas seed oil contents in 2010-2011 were higher than that in 2009-2010. The highest yield appeared in CHB, which was significantly higher than that in MH. Furthermore, harvesting loss in CHB were 50% that in MH. Seed oil content and chlorophyll exhibited no obvious difference between CHB and MH. Economic profit analysis demonstrated that mechanical sowing/combine harvesting （MS/CH） showed an input/output ratio of 1：1.6, and it was 1：1.2 in mechanical sowing/manual harvesting （MS/MH）. Labor-cost accounted for more than 70% of the total cost in MS/MH, which led to low profitability to a great extent. Our results suggested that CHB was the optimum harvesting time for winter oilseed rape along the Yangtze River.

Nutrient deficiency limits population development,yield formation,and nutrient uptake of direct sown winter oilseed rape

Direct-sowing establishment method has great significance in improving winter oilseed rape（Brassica napus L.） production and guaranteeing edible oil security in China. However, nutrient responses on direct sown winter oilseed rape（DOR） performance and population development dynamic are still not well understood. Therefore, five on-farm experiments were conducted in the reaches of the Yangtze River（RYR） to determine the effects of nitrogen（N）, phosphorus（P）, and potassium（K） deficiencies on population density, dry matter production, nutrient uptake, seed yield, and yield components of DOR plants. Four fertilization treatments included the balanced NPK application treatment（NPK, 180 kg N, 39.3 kg P, 100 kg K, and 1.8 kg borax ha–1） and three nutrient deficiency treatments based on the NPK treatment, i.e., –N, –P, and –K. The results indicated that DOR population density declined gradually throughout the growing season, especially at over-wintering and pod-development stages. Nutrient deficiency decreased nutrient concentration in DOR plants, limited dry matter production and nutrient uptake, and thereby exacerbated density reduction during plants growth. The poor individual growth and reduced population density together decreased seed yield in the nutrient deficiency treatment. Averaged across all the experiments, seed yield reduced 61% by N deficiency, 38.3% by P deficiency, and 14.4% by K deficiency. The negative effects of nutrient deficiency on DOR performances followed the order of –N–P–K, and the effects were various among different nutrient deficiencies. Although N deficiency improved DOR emergence, but it seriously limited dry matter production and nutrient uptake, which in turn led to substantial plants death and therefore resulted in a very low harvested density. The P deficiency significantly reduced initial density, limited plants growth, and exacerbated density reduction. The K deficiency mainly decreased individual growth and yield, but did not affect density dynamic. Our results highlighted the importance of balanced NPK application in DOR production, suggesting that management strategy of these nutrients should be comprehensively considered with an aim to build an appropriate population structure with balanced plant density and individual growth.

油蔬两用油菜品种的品质与产量评价

为促进油菜油蔬两用技术的推广,对18个油蔬两用油菜品种的菜薹产量、不摘薹和摘薹后菜籽产量等性状,以及菜薹的粗纤维、可溶性糖、维生素C、钙、铁、锌、硒质量分数和食味得分等品质性状进行统计,开展主成分分析和聚类分析,并计算各品种的综合产值.结果表明:11个性状可综合为6个主成分,18个品种可划分为3个类群;基于产量和品质特征确定了3个总产值高的品种赣油杂8号、邡油777和沣油958以及1个优质高产品种渝南油683,结果可为重庆地区油菜油蔬两用技术的推广提供种质基础和数据支撑.

鄂中地区油菜专用肥施用效果

为明确油菜(Brassica napus L.)专用肥对鄂中地区直播油菜产量的提升效果,于2019—2021年在湖北省沙洋县开展了连续2年的田间试验,设置了不施肥、常规施肥、宜施壮专用肥和洋丰专用肥4个施肥处理,考察各处理下成熟期油菜产量、地上部生物量、地上部养分含量、地上部养分积累量以及肥料偏生产力等的差异。结果表明,与不施肥对照相比,常规施肥和专用肥处理均能显著提高油菜子产量;与常规施肥相比,专用肥处理总养分投入减少了41.3%,而两者油菜子产量无明显差异。施肥显著增加了油菜地上部生物量,促进了油菜地上部对养分的吸收。油菜专用肥处理显著提高了肥料偏生产力,与常规施肥处理相比,宜施壮专用肥和洋丰专用肥处理平均肥料偏生产力在2019—2020年、2020—2021年分别提高了80.6%和68.2%。综上所述,油菜专用肥在保障油菜高产稳产的同时,提高了肥料利用率。

甘蓝型杂交油菜新品种油蔬两用潜力分析

为了分析甘蓝型油菜在川南地区的油蔬两用潜力,选用6个甘蓝型油菜品种和2个蔬菜品种,从经济效益、农艺性状和营养品质3个方面进行分析。结果表明,油蔬两用种植技术虽然降低了菜籽产量,但是能增加经济效益。摘薹处理损失了主茎和主茎角果,同时减少了单株一次分枝数和一次分枝角果数,进而间接影响每角粒数,导致菜籽减产。甘蓝型油菜菜薹是一种适口性好、营养价值丰富的功能型蔬菜。该研究证实了油蔬两用是一项适宜推广的种植技术。

Identification of QTLs for Branch Number in Oilseed Rape(Brassica napus L.)

Oilseed rape （Brassica napus L.） is one of the most important oil crops worldwide and provides about 50 percent of the vegetable oil supply in China （Yin et al., 2009）. The development of rapeseed varieties with higher yield is an effective measure to optimize balance between the supply and demand of edible vegetable oil. In oilseed rape, the number of silique per plant （SP） contributes most to the yield performance （Diepenbrock, 2000）. However, compared with the other two yield-component traits, seeds per silique （SS） and seed weight （SW）, SP is more sensitive to environmental changes （Li et al., 2007; Shi et al., 2009）. Therefore, it is difficult to perform the genetic improvement on SP trait directly in oilseed rape.

Reaction of Oilseed Rape Cultivars to Sclerotinia Stem Rot in Field Environ- ments

Sclerotinia sclerotiorum (Lib.) de Bary, as an ubiquitous phytopathogenic Ascomycete fungus capable of infecting a wide range of plants, has increased in importance for oilseed rape production worldwide in recent years. The objective of this research was to determine the effectiveness of two measurements of disease rating in predicting the field reaction of cultivars to sclerotinia stem rot. One hundred and ten cultivars or lines of oilseed rape were repeatedly evaluated the disease severity index (DSI) and lesion length for their resistance to nine isolates of S. sclerotiorum in two field stations. There were greatly significant differences between two stations, among isolates, among population of cultivars for the disease severity index and lesion length, and also significant station×isolate interaction for lesion length and population×isolate interactions for DSI and lesion length, but interactions of station×isolate for DSI were not significant. Cultivars such as Pomorzanin and Lisek consistently had the lowest disease severity ratings in the field tests. However, most of cultivars were susceptible to sclerotinia stem rot in the field test. A positive correlation between the mean DSI and lesion length was found for both isolates (Sc23 and B1). The correlations ranged from 0.23 to 0.83 at various plots, with nine of the twelve correlations significant. The correlations for disease caused by different fungal isolates were statistically significant difference. These correlations ranged from -0.44 to 0.90, with nine of the thirty-two correlations significant. It indicated that resistance to some isolates was significantly correlated and selection of some varieties for resistance to one isolate appeared to improve resistance to other isolates.

Effects of harvesting method and date on yield loss and seed quality of rapeseed

China is one of the major rapeseed production countries in the world,but harvesting mechanization was still backward,and high harvest loss was a key inhibiting factor for rapeseed production.To obtain optimum harvesting date for winter rape in the Yangtze River Valley of China,artificial simulated combine harvesting and artificial two-stage harvesting were adopted to find correlations between harvesting date,rape variety,seed oil content,protein content,grain moisture rate,rapeseed straw moisture content,rapeseed unthreshing rate,grain drop loss rate,thousand seeds weight(TSW),and harvesting economic coefficient(HEC).Analysis of variance showed that rapeseed oil content,protein content and TSW were correlated with rape variety;HEC was correlated with harvesting method and rape variety.Rape variety was the dominant factor of rapeseed oil content,protein content,TSW and HEC.Grain moisture rate,un-threshing rate,grain drop loss rate and straw moisture content were correlated with harvesting method and date.Harvesting date was the dominant factor of rapeseed moisture rate,un-threshing rate,grain drop loss rate and straw moisture rate.Single factor tests further proved that harvesting date had less impact on rapeseed oil content,protein content,TSW and HEC.It showed little correlation with rapeseed quality except grain moisture rate.The optimum harvesting date could be known by change in straw moisture content.The combined harvesting should be carried out during the grain moisture content of 15%-20%with TSW stabilized in the highest level.The twostage harvesting cutting should be carried out at the grain moisture content of 35%-40%.This research offered a reference to harvesting method and date for rape cultivated in the Yangtze River Valley.

氮肥用量对冬油菜籽粒产量和品质的影响

为探明不同氮肥用量对冬油菜籽粒产量和品质的影响,于2019/2020以及2020/2021年在湖北省武穴市开展田间试验,试验设置0、90、180、270、360 kg N hm^(-2)5个氮肥施用水平。在成熟期测定油菜籽产量、氮含量和油菜籽品质指标。结果表明,施氮显著提高冬油菜籽粒产量及氮含量,与不施氮相比,氮肥施用后平均增产1548 kg hm^(-2),平均增产率达32.9%,在施氮量为0~270 kg N hm^(-2)范围内,油菜籽产量随氮肥施用量显著增加,继续增施氮肥,油菜籽产量无明显变化或有下降趋势,施氮主要通过提高单株角果数来提高油菜籽粒产量。施氮显著增加了籽粒蛋白质含量,当施氮量达270 kg N hm^(-2)时籽粒氮含量和蛋白质含量最高。施氮显著降低籽粒含油量,氮肥用量每增加100 kg N hm^(-2),籽粒含油量下降1.6个百分点。随着氮肥用量的增加,籽粒硫甙、油酸、亚麻酸、芥酸以及饱和脂肪酸(棕榈酸和硬脂酸)呈升高趋势,亚油酸呈降低趋势,油菜籽粒品质整体呈降低趋势。综上所述,以追求高品质食用油为目标,同时兼顾油菜籽粒产量,氮肥适宜用量约为180kgNhm^(-2);以追求油菜籽粒产量并考虑油菜饼粕作饲料为目标时,氮肥适宜用量约为270 kg N hm^(-2)。

不同轮作模式下氮肥施用对油菜产量形成及养分利用的影响

【目的】探究施氮量对不同轮作模式下油菜产量形成及养分利用的影响。【方法】以甘蓝型油菜品种‘中油杂19’为材料,在湖北黄冈进行大田裂区试验,设置两个轮作模式(稻油轮作,RO;豆油轮作,SO)为主区,4个施氮量(N0,0;N1,90 kg·hm^(-2);N2,180 kg·hm^(-2);N3,270 kg·hm^(-2))为副区,测定产量构成、干物质积累、农艺性状、养分积累及籽粒品质等相关指标。【结果】(1)豆油轮作油菜籽粒产量显著高于稻油轮作,增加施氮量,不同轮作模式下油菜单株角果数、每角果粒数及千粒重均呈显著上升趋势,与N0相比,在N1、N2和N3处理下,稻油轮作油菜籽粒产量分别增长176.68%、436.49%和835.40%,豆油轮作油菜籽粒产量分别增长123.96%、344.46%和547.25%。豆油轮作较稻油轮作在N0、N1、N2和N3处理下大田籽粒产量分别增长62.09%、31.33%、71.79%和12.21%;(2)成熟期豆油轮作油菜根颈粗、株高、第一有效分枝高度和有效分枝数显著高于稻油轮作,且随施氮量增加不同轮作模式下各农艺性状指标显著增加;各生育期豆油轮作单株油菜根干重及地上部干重显著高于稻油轮作,但根冠比低于稻油轮作,且随施氮量增加,两种轮作模式下苗期后根冠比显著下降;(3)豆油轮作油菜根系、角果壳、茎秆、籽粒氮含量和氮积累量均高于稻油轮作,且随着施氮量增加各部位氮含量和氮积累量显著增加。豆油轮作籽粒氮素表观利用率高于稻油轮作,随施氮量的增加,稻油轮作下氮素表观利用率增加,而豆油轮作氮素表观利用率呈现先上升后下降趋势;(4)与稻油轮作模式相比,相同施氮量下豆油轮作油菜角果壳可溶性糖含量低,而游离氨基酸含量和游离氨基酸含量/可溶性糖含量比值高,随施氮量增加可溶性糖含量降低,游离氨基酸含量和游离氨基酸含量/可溶性糖含量比值增加。因此,豆油轮作油菜籽粒因脂肪酸合成底物受限,含油率低于稻油轮作模式,且随施氮量增加,籽粒含油率在各轮作模式下均显著下降。两年试验结果显示,在270 kg·hm^(-2)施氮量水平时,产油量在两个轮作模式下均达到最大,稻油轮作两年产油量分别为1678.60和1665.33 kg·hm^(-2),豆油轮作两年产油量分别为1684.03和1687.10 kg·hm^(-2),但豆油轮作在180和270 kg·hm^(-2)施氮量下产油量差异不显著。【结论】稻油轮作油菜氮肥施用可控制在270 kg·hm^(-2)左右,而豆油轮作氮肥施用可控制在180 kg·hm^(-2)左右,以保证较高氮肥利用效率,并获得较高产油量。

采薹对甘蓝型油菜农艺性状和经济效益的影响

为了提高油菜应用价值,探索油菜多功能利用新思路。选择农艺性状优异的甘蓝型油菜G1为父本,G2为母本,构建289个双单倍体(DH)纯系,对其菜薹产量、植株性状、菜籽产量、生物学产量与经济效益进行考察评估,筛选菜薹产量高,不影响菜籽产量且经济效益高的“油薹两用”油菜品系。结果表明:分枝高度(BH)、一次有效分枝数(NFB)与菜籽产量呈负相关关系;与对照相比,采摘菜薹降低了株高(PH)、分枝高度和一次有效分枝数,差异达到显著水平;DH群体中,菜薹产量与菜籽产量呈负相关关系;摘薹后菜籽产量比未摘薹平均减少19.24%,但总产值比未摘薹平均增加48.87%,其中筛选出48份材料菜籽产量(产量变化范围在-10%~10%)不受摘薹影响,63份材料摘薹后菜籽产量增加,增产幅度>10%,增产效果明显。30份材料菜籽产量超过3000 kg/hm^(2)。综合以上指标,本试验共筛选出21份菜薹与菜籽产量均达到长江流域油菜区产量要求且效益好的“油薹两用”材料,为兼用型油菜的选育及推广提供理论指导和科学依据。

肇庆市油菜引种比较研究

[目的]为了推进肇庆市的油菜种植,选取8个甘蓝型油菜品系进行引种试验。[方法]设置2个不同播期,采用随机区组设计,分别考察了各品系的长势、虫害情况、生育期以及终花期株型相关农艺性状,同时比较了第2播期的水淹条件对不同品系各项性状的抑制作用。[结果]F品系花期最早,只需要53 d时间,但容易倒伏,而G品系的株高最高约188 cm,E品系的开展度最大接近80 cm,极易倒伏。水淹条件缩短了各品系的开花期,显著抑制了各品系的株高和开展度。相比之下,D品系耐淹能力最强,株高和开展度分别减少39和5 cm,冠层高度反而增加13 cm。[结论]综合考虑肇庆市多功能油菜产业对早熟且耐水淹油菜品系的需求,下一步仍需要对引种材料进行杂交改良。

油菜的遗传转化及抗溴苯腈转基因油菜的获得

以油菜(Brassica napus L.)的下胚轴和子叶为转化受体,建立了油菜的高效转化系统。在此基础上,将抗除草剂溴苯腈基因(bxn基因)导入油菜,获得了抗溴苯腈转基因油菜。分子检测实验证明,转基因油菜中含有bxn基因。转基因油菜可抗高达10^(-3)mol/L的溴苯腈。

甘蓝型双低油菜油蔬两用栽培的产量、效益及菜薹营养研究初报

以甘蓝型双低油菜和白菜型红菜薹为材料,通过对比试验和化学检测研究了甘蓝型双低油菜油蔬两用栽培在重庆市生态生产条件下的产量、效益和菜薹营养。结果表明:油蔬两用栽培产菜薹2382 kg/hm2,产菜籽2219 kg/hm2,菜籽产量比单收菜籽栽培减产8.1%;油蔬两用栽培纯收入5221.80元/hm2,比单收菜籽栽培新增纯收入1552.80元/hm2,增收42.3%;甘蓝型双低油菜薹营养丰富,除铁的含量外,其维生素C和钙、硒、锌等微量元素含量明显高于白菜型红菜薹,可溶性总糖、粗纤维等营养指标与白菜型红菜薹相当。甘蓝型双低油菜油蔬两用栽培有良好的增收效果,可在重庆市适宜地区示范推广。

不同品种油菜子粒产量及氮效率差异研究

采用大田试验,以16个冬油菜品种为试验材料,系统研究了油菜子粒产量、氮素吸收量、氮素响应度和氮素利用效率的品种间差异,并初步探讨了氮素吸收效率和氮素利用效率对不同品种油菜氮效率差异的贡献。结果表明,无论施氮水平如何,不同品种的子粒产量、氮素利用效率和氮素响应度均有显著差异,而氮素吸收量只有在不施氮条件下品种间差异才达到显著水平。根据不施氮时的氮效率和氮素响应度将16个油菜品种分为4种不同类型:1)氮高效-高氮响应(NHE-NHR)型,包括Xy1、Xy16、Xy17、Xh19、Xh20和Xy21;2)氮低效-低氮响应(NLE-NLR)型,包括Xy6、Xy8和Xy9;3)氮高效-低氮响应(NHE-NLR)型,包括Xy7、Xy12、Xy14、Xy15和Xy24;4)氮低效-高氮响应(NLE-NHR)型,包括Xy11和Xy13。无论供氮水平如何,氮素利用效率的变异系数均大于氮素吸收效率的变异系数,说明氮素利用效率对油菜氮效率差异的贡献大于氮素吸收效率。但是,氮素吸收效率的变异系数表现为不施氮时大于施氮,氮素利用效率的变异系数则相反,说明在氮胁迫条件下,氮效率的差异中来源于氮素利用效率的变异减少,来源于氮素吸收效率的变异增加。

播期及种植密度对直播油菜农艺性状和产量品质的影响

本文于2010-2011年在四川雅安（盆周山区）、射洪（川中丘陵区）和邛崃（成都平原区）3个不同试验地点,研究了播种期、种植密度对直播油菜农艺性状和产量品质的影响。结果表明,直播油菜不同试验地点的农艺性状、产量和品质存在着一定的差异。播期是影响直播油菜株高、一次分枝数和产量的主要因素;种植密度则是影响直播油菜分枝部位的主要因素。株高、分枝部位和一次分枝数均随播期推迟而降低或减少;株高和一次分枝数则随种植密度增加而降低。直播油菜籽粒产量和油产量都随播期推迟而呈降低趋势,但油菜产量在不同种植密度下并无显著差异。播期、种植密度对直播油菜籽粒含油量、油酸含量和亚油酸含量均无显著性影响。基于上述结果,对于四川盆地不同生态区的直播油菜,在目前收获机械难以进入的深丘山区稻油两熟制农田,可适期早播（9月下旬）,种植密度以每公顷15~30万株为宜;在四川省有条件发展油菜机械化生产的浅丘平坝区,适宜播期（9月下旬至10月中旬）范围内,随播期推迟适当加大种植密度（每公顷30~60万株）,通过构建小个体大群体结构,从而实现油菜生产的轻简高效。