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

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

Transcriptional proffling between yellow-and black-seeded Brassica napus reveals molecular modulations on flavonoid and fatty acid content

Brassica napus is an important cash crop broadly grown for the vegetable and oil values.Yellow-seeded B.napus is preferred by breeders due to its improved oil and protein quality,less pigments and lignin compared with the blackseeded counterpart.This study compared the differences in flavonoid and fatty acid contents between yellow rapeseed from the progenies of B.napus-Sinapis alba somatic hybrids and the black-seeded counterpart using RNA-seq analysis.Through HPLC-PDA-ESI(-)/MSanalysis,it was found that phenylpropanoids and flavonoids(i.e.,isorhamnetin,epicatechin,kaempferol,and other derivatives)in yellow seed were significantly lower than those in black seed.The fatty acid(FA)content in yellow rapeseed was higher than that in black rapeseed due to the variation of C16:0,C18:0,C18:1,C18:2,and C18:3 contents.RNA-seq analysis of seeds at four and five weeks after flowering(WAF)indicated that differentially expressed genes(DEGs)between black and yellow rapeseeds were enriched in flavonoid and FA biosynthesis,including BnTT3,BnTT4,BnTT18,and BnFAD2.Also,genes related to FA biosynthesis,desaturation and elongation(FAD3,LEC1,FUS3,and LPAT2)in yellow seed were up-regulated compared to those in black seed,while genes involved in beta-oxidation cycle(AIM1 and KAT2)of yellow seed were down-regulated compared to those in black seed.The DEGs related to the variation of flavonoids,phenylpropanoids,and FAs would help improve the knowledge of yellow seed character in B.napus and promote rapeseed improvement.

Genetic Diversity of Testa Pigments and RAPD Marker of Yellow-Seeded Rapeseed (Brassica napus L.)

14 yellow-seeded rapeseed lines (Brassia napus L.) from different genetic sources were used to analyze diversity of testa pigments content, oil and protein content, and RAPD markers. The results showed that the anthocyanin and melanin were the most important pigments in testa and their content were responsible for the variation in seed color ranging from orange to black yellow, 14 yellow-seeded lines could be classified into 3 groups: high anthocyanin content group with anthocyanin content over 2. 54 mg g-1 DW, the seed color was light yellow or orange; low pigments content group with low content of anthocyanin and melanin, the testa was transparent and the seed color was light yellow, greenish yellow or twany; high melanin content group with melanin content over 178. 4U(A290nm) , the testa was black, the seed color was black yellow. Oil content changed from 36.2% to 45. 5%, protein content from 21.1% to 27.7% , and the correlation analysis revealed that the oil content is highly significantly negatively correlated with the protein content. The cluster analysis showed that the extensive genetic variation existed among 14 yellow-seeded lines by using unweighted paired group method with arithmetic average (UPGMA) based on RAPD markers which were amplified with decamer primers, the genetic similarity among them ranged from 0. 25 to 0.909, and 14 yellow-seeded lines could put into 2 clusters corresponding to genome difference.

Physiological Differences Between Yellow-Seeded and Black-Seeded Rapeseeds (Brassica napus L.) with Different Testa Characteristics During Artificial Ageing

Yellow-seeded rapeseed (Brassica napus L.) is a new kind of breeding resources with yellow color, increased oil and protein content and less unwanted crude fiber content due to the thinner and transparent testa compared with traditional black or brown-seeded rapeseed. To analyze the longevity of the yellow-seeded rapeseed during storage, the physiological differences between the yellow and black-seeded near-isogenic lines were studied by artificial ageing method. The testa rate, anthocyanin content and melanin content of yellow-seeded rapeseeds decreased by 20.1, 25.2, and 80.4% respectively than black-seeded rapeseed. During artificial ageing, the yellow-seeded rapeseed showed significantly different effect of ageing compared with the black-seeded, as demonstrated by faster deterioration with lower germination percentage, seed vigour index, reducing sugar and soluble protein contents than the black-seeded, as well as a drastic increase in electrical conductivity, malnodialdehyde (MDA) content and a rapid decrease in activities of antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD). The results suggested that the transparent testa of the yellow-seeded rapeseed lost some abilities to protect the embryo against adverse environmental conditions and thus led to a poor storability.

Study on Seedling Techniques of Artocarpus nanchuanensis

[Objective] This study aimed to solve the problems of low seed germina- tion rate and difficult bareroot seedling transplanting in artificial cultivation of Arto- carpus nanchuanensis, a native species in China. [Method] Under different seed treatment and seedling style, the seed germination rate and seedling emergence rate of A. nanchuanensis were investigated. [Result] tn both sand bed and direct sowing yellow soil, the seed germination rate, seedling survival rate and seedling emergence rate in the treatment II were all higher than those in the treatments I and III. The average seed germination rate, seedling survival rate and seedling emergence rate in sand beds were 1.5%, 16.27% and 13.95%, respectively higher than those in direct sowing yellow soil. The average seedling emergence rate in sand beds was 2.46% higher than that in containers. Among the 8 kinds of light matrix formulas, the growth of seedlings in the formula VIII was best. In the three treatment groups, the seedling emergence rates, seedling heights and ground diame- ters in the formula VIII were all significantly different from those in the other formu- las. The seedling growth in the formula I and III ranked second, while the seedling emergence rate, seedling height and ground diameter in the formula VII were the poorest. [Conclusion] If conditions allow, the formulas I （meadow soil：vermiculite：yel- low soi1=5：3：2）, III （meadow soil：perlite：yellow soi1=5：3：2） and VIII （peat：forest top soi1=6：4） should be promoted in seedling with light matrixes.

Green Control Measures of Weeds in Wheat Fields in Hetao Irrigation Area along the Yellow River

This study was conducted to solve the problem of green weed control in wheat fields in Hetao irrigation area among the Yellow River.Based on the observation of the competition between wheat and weeds in areas where weeds occurred seriously in wheat fields in Hetao irrigation area among the Yellow River,we measured the effects of green weed control measures and wheat yield using different wheat varieties,planting densities,different organic fertilizers,different ploughing times,and different mulching methods.The results showed that the emergence of weeds in wheat fields dominated by Chenopodiaceae weeds,grain amaranth and barnyard grass was more than 10 d later than wheat.Weeds were mainly distributed between rows(holes),and the number of plants accounted for 66.6%(drill seeding)and 97.6%(hole seeding),respectively.And the growth of weeds in rows(holes)was weaker,and the fresh weight of individual plants was 39.3%-41.9%lower than that between rows(holes).The ecological weed inhibitory effect was significant in the early stage of wheat growth;and among the green weed control measures,except that different varieties and planting densities caused no significant difference in weed control effect,other measures had obvious weed control effects.Comprehensive comparison showed that the control effects of plant number in black film full-covered hole seeding,conventional film-covered hole seeding,increasing ploughing times,and applying organic fertilizer free of weed seed pollution were 82.3%,71.7%,22.0%,and 8.6%,respectively;the fresh weight control effects of black film full-covered hole seeding,conventional film-covered hole seeding,increasing ploughing times,and applying organic fertilizer free of weed seed pollution were 98.0%,97.1%,23.9%,and 9.6%,respectively;and the fresh weight control effects of black film full-covered hole seeding,conventional film-covered hole seeding and increasing ploughing times increased wheat yield by 69.4%,56.4%and 21.1%,respectively.The technologies in this study can realize the purposes of mechanized green weed control in organic wheat production and low-cost,high-yield,large-scale production.

促生菌黄浆水培养基优化及发酵菌液对种子萌发的影响

黄浆水含有大量的营养物质,对其资源再利用具有重要意义。通过水解圈的定性分析及氨氮含量的定量测定以筛选具氨化及溶磷作用的促生菌菌株,通过单因素、正交试验对2个菌株的黄浆水发酵培养基进行优化,利用2个菌株的发酵菌液对黑麦草及苜蓿种子进行浸种以分析其对种子萌发的影响。结果表明:恶臭假单胞菌(Pseudomonas putida)HGD3和吡咯伯克霍尔德氏菌(Burkholderia pyrrocinia) P10具有较强的氨化能力和溶磷性。在含40%黄浆水的优化发酵培养基中培养48 h, HGD3和P10菌株发酵液的氨氮浓度分别为894.92 mg/L和546.49 mg/L,较优化前分别提高6.79和2.60倍。HGD3和P10菌株发酵液的浸种均显著地提高了黑麦草和苜蓿种子的发芽率,苜蓿种子的发芽率较对照分别提高了34.51%和24.36%;黑麦草种子的发芽率分别提高了32.66%和18.85%,且萌发黑麦草的芽长分别增长1.57倍和0.77倍、根长分别增长了1.53倍和0.91倍。恶臭假单胞菌HGD3的黄浆水发酵液可以显著地促进黑麦草和苜蓿种子的萌发,尤以黑麦草为甚,研究结果为黄浆水的再利用及促生菌菌株对种子的促萌发提供了一定的理论基础。

不同种植密度与氮肥互作对红花产量及品质的影响

为明确引黄灌区红花(Carthamus tinctorius)最佳种植密度和适宜氮肥施用量,本试验选用红花‘甘红1号’为试验材料,采用裂区设计,主区为施氮量,副区为种植密度,设3个施N水平(45、90、135 kg·hm^(-2))和4个种植密度(株行距分别为30 cm×60 cm、25 cm×60 cm、25 cm×50 cm、27 cm×40 cm),研究不同施氮量和种植密度对红花产量品质及氮素吸收利用的影响。结果表明,施氮量和种植密度显著影响红花产量品质及氮素利用,农艺性状、产量性状及氮素积累量随着两者的增加呈先增大后降低的趋势。籽粒产量、羟基红花黄色素A (HSYA)、氮素积累量、氮肥表观利用率和氮肥农学利用率受交互作用影响显著,干花产量对种植密度反应敏感。其中,施氮量90 kg·hm^(-2)、株行距25 cm×50 cm条件下,HSYA含量最高,其余指标在施氮量90 kg·hm^(-2)、株行距25 cm×60 cm条件下趋于最大。综合考虑产量、品质、氮素利用协同提高,在引黄灌区红花种植最佳施氮量为90 kg·hm^(-2),种植密度以株行距25 cm×50 cm或25 cm×60 cm为宜。

智能播种机器人结构设计与试验

针对当前黄河流域治沙绿化中地形复杂、人工种植效果差、大型设备能源结构不合理等问题,设计了一种集图像传输、远程控制和光伏发电系统等功能于一体的智能播种机器人。首先进行了播种机器人的总体方案设计;然后根据播种机器人的工作区域特点对关键零部件进行了计算和选型,利用SolidWorks软件对各零件进行建模及装配,并应用Simulation软件对关键零部件进行了静应力分析和模态分析;最后对该播种机器人进行样机制作,播种试验结果表明装置的播种合格率达到86.08%。此播种机器人的推广可以降低劳动者的工作强度,对提高农业生产的智能化具有重要意义。

不同光质对吉定黄籽油菜生长发育影响初探

为研究不同光质对西藏白菜型黄籽油菜生长发育的影响,以西藏地方品种,吉定黄籽油菜为实验材料,播种于盛有有机质、腐殖酸混合基质的花盆中,并置于7种不同的光质下进行培养:荧光灯(FL)、LEDs蓝光(B)、LEDs红光(R)、LEDs白光(W)、LEDs绿光(G)以及不同LEDs红、蓝配比光质(RB2:1;3:1)。结果表明,LEDs绿光处理,油菜株高最高,为5.95 cm,荧光灯处理株高为4.75 cm,属最低;单色光(红光、绿光、蓝光、白光)处理的幼苗高度、生长速率、根长、株高等指标均大于在复合光质处理的幼苗;而干鲜重、叶绿素的含量、蛋白质含量、叶面积等指标复合光质处理下高于单色光处理的油菜幼苗。由此可见,油菜营养物质的合成与积累与光质有关,且单色光不适合油菜幼苗的生长。

不同遗传背景的甘蓝型黄籽油菜粒色遗传初步研究

以７种不同来源的甘蓝型黄籽油菜品系为材料，采用完全双列杂交试验和与中油８２１测交的方法，分析了粒色的遗传规律。初步结果显示：不同来源的甘蓝型黄籽油菜品系带有不同的粒色基因，有些为等位基因，有些属非等位基因。粒色遗传以加性效应为主，显性和上位性效应也很明显。

缺氧对甘蓝型黄籽与黑籽油菜发芽的影响

随机选取甘蓝型黄籽、黑籽油菜各20个品系,将萌发中的种子进行淹水缺氧处理,观察黄籽、黑籽品系耐湿性表现。结果表明,甘蓝型黄籽油菜的相对成苗率、相对苗高、相对苗重、活力指数低于黑籽油菜,单位电导率高于黑籽油菜。无论是黄籽油菜还是黑籽油菜,品种间的耐湿性差异都很大。电导率与活力指数之间有极显著的负相关,可用来快速筛选油菜耐湿性材料。

氮磷钾硼对甘蓝型黄籽油菜产量和品质的影响

采用四元二次回归正交旋转组合设计,以氮(N)、磷(P)、钾(K)、硼(B)肥为主要探讨因子进行田间试验,建立N、P、K、B4因素与黄籽油菜籽粒产量、含油量、产油量以及蛋白质含量的施肥模型。对模型解析发现,在供试条件下N、P、K、B单因子对产量和产油量的影响均是X1(N)>X2(P2O5)>X4(B)>X3(K2O),对含油量的影响是X1(N)>X4(B)>X2(P2O5)>X3(K2O),对蛋白质的影响为X1(N)>X4(B)>X3(K2O)>X2(P2O5)。施肥模型寻优结果表明施N163.7～179.2kghm-2、P2O585.0～95.0kghm-2、K2O100.7～124.3kghm-2、B6.7～8.3kghm-2,可使黄籽油菜产油量达到1000kghm-2,饼粕蛋白质含量达40%以上;饼粕蛋白质含量大于45%时,各养分因子的取值区域分别是施N236.6～255.4kghm-2、P2O580.3～99.7kghm-2、KO103.2～126.8kghm-2、B8.9～10.3kghm-2。

利用SSR标记对甘蓝型黄籽油菜贵油519杂种纯度鉴定

在用SSR(简单重复系列)标记构建了53份甘蓝型黄籽油菜种质资源的DNA指纹图谱的基础上,筛选出一个对黄籽油菜杂种贵油519及亲本具有共显性的SSR引物Ra2A05b,鉴定大田生产F1杂种群体的纯度,并对比田间调查的鉴定结果表明,SSR标记鉴定可以识别更多的非杂交种单株,如因微粉导致的自交、花粉污染、机械混杂等带来的非杂交种单株。就SSR标记鉴定杂交种纯度鉴定的可行性和优点进行了讨论。

培养条件对甘蓝型黄籽油菜下胚轴的再生影响

以甘蓝型黄籽油菜GH01的下胚轴为材料,研究影响外植体芽再生的因素。试验结果表明,苗龄、预培养基的激素浓度和预培养时间、分化培养基等对芽再生频率均有较大影响。8d苗龄的下胚轴置 MS+1.5mg/L 2,4-D+0.1mg/L 6-BA培养基预培养4d后,转分化培养基MS+4.0mg/L 6-BA+0.05mg/L NAA+5.0mg/L AgNO_3+0.6mg/L GA_3 培养,可获得较高的芽再生频率。添加 5mng/L AgNO_3 处理可防止外植体褐化并有利于芽分化;0.6mg/L GA_3 处理可促进胚状体形成,提高芽再生频率。GH01 最高芽再生频率为40.50％。

甘蓝型黄籽油菜主要性状的遗传相关分析

采用混合线性模型分析方法,以7个不同遗传来源的甘蓝型黄籽品系为材料配制完全双列杂交,用ADM模型对甘蓝型黄籽油菜主要性状进行遗传相关分析。结果表明,主序有效角果数与单株有效角果数以显性相关为主,主要体现在杂种一代。单株有效角果数与千粒重、千粒重与单株粒重在遗传上没有紧密的联系。每角粒数、一次有效分枝数、单株有效角果数、角果长度、株高。纤维素含量和黄籽度等是影响单株粒重的主要遗传因子,通过单个性状间接选择不能有效改良单株粒重。

黄籽羽衣甘蓝和白菜型油菜杂交再合成甘蓝型油菜研究

为了将携带有黄籽基因的芸薹属C和A染色体组结合在一起,开展了黄籽羽衣甘蓝和黄籽白菜型油菜种间杂交再合成甘蓝型油菜研究。在MS+1 0mg/LBA+0 2mg/LNAA+7%蔗糖+300mg/LCH培养基上培养授粉10d后的40个子房获得2个白菜型油菜×羽衣甘蓝杂种,其中1个自然加倍成为双二倍体。再合成甘蓝型油菜双二倍体和单倍体分别结黑籽和红褐籽。

甘蓝型黄籽油菜主要性状的通径分析

以7个不同遗传来源的甘蓝型黄籽品系配制的完全双列杂交组合作为样本,对甘蓝型黄籽油菜主要性状进行通径分析。主要结果为:全株有效果数和株高对单株粒重有最大正向作用,通径系数分别为0.514和0.535。皮壳率对黄籽度有直接的最大负效应,通径系数为-0.798。

甘蓝型黄籽油菜SSR标记遗传多样性

利用 42对SSR引物对19个甘蓝型黄籽油菜品种(系)进行遗传多样性分析,共检测出336个等位基因,每对引物在不同品系间的等位基因数在2～21之间,平均位点为8个.位点多态性信息含量为0.17～0.85,平均为0.73.对SSR扩增结果进行UPGMA分析,可将19个品种(系)分为4个群体,聚类结果与系谱来源比较一致.并且只需NAS99、NAS98、NAS91及NAS31 4对多态性高的引物即可将19个材料区分开来.

不同施肥水平对甘蓝型黄籽油菜含油量的效应研究

采用4因素正交旋转组合设计,研究了氮、磷、钾、硼施用量对供试品种渝黄1号和GH06种子含油量的影响。结果表明:1)氮、磷、钾、硼养分因子对含油量均有较大影响;2)施用氮肥有降低含油量的作用,而钾、硼肥有提高含油量的作用,增施磷肥在一定范围内可以提高含油量,超出适宜范围则会降低含油量;3)氮、磷肥水平之间有较显著的互作效应,并与土壤本身的理化性状有关。