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.

Synthesis and characterization of yellow water-borne polyurethane using a diol colorant as extender

A novel and facile method toward a series of yellow water-borne polyurethane was developed by using an intrinsically colored diol in this paper.The yellow aqueous dispersion PCLD-HENA-PU was synthesized based on isophorone diisocyanate(IPDI), polycaprolactonediol(PCLD) and 2,2-dimethylol propionic acid(DMPA) using a yellow diol N,N-bis(2-hydroxyethyl)-4-nitroaniline (HENA) as a chain extender.Due to the complete reaction of OH group in colorant HENA with NCO group in diisocyanate,a series of stable yellow p...

Identification of a novel seed size associated locus SW9-1 in soybean

Seed size is one of the vital traits determining seed appearance, quality, and yield. Untangling the genetic mechanisms regulating soybean 100-seed weight (100-SW), seed length and seed width across environments may provide a theoretical basis for improving seed yield. However, there are few reports related to QTL mapping of 100-SW across multiple ecological regions. In this study, 21 loci associated with seed size traits were identified using a genome-wide association of 5361 single nucleotide polymorphisms (SNPs) across three ecoregions in China, which could explain 8.12%–14.25% of the phenotypic variance respectively. A new locus, named as SW9-1 on chromosome 9 that explained 10.05%–10.93% of the seed weight variance was found significantly related to seed size traits, and was not previously reported. The selection effect analysis showed that SW9-1 locus has a relatively high phenotypic effect (13.67) on 100-SW, with a greater contribution by the accessions with bigger seeds (3.69) than the accessions with small seeds (1.66). Increases in seed weight were accompanied by increases in the frequency of SW9-1T allele, with >90% of the bred varieties with a 100-SW >30 g carrying SW9-1T. Analysis of SW9-1 allelic variation in additional soybean accessions showed that SW9-1T allele accounting for 13.83% of the wild accessions, while in 46.55% and 51.57% of the landraces and bred accessions, respectively, this results indicating that the SW9-1 locus has been subjected to artificial selection during the early stages of soybean breeding, especially the utilization of SW9-1T in edamame for big seed. These results suggest that SW9-1 is a novel and reliable locus associated with seed size traits, and might have an important implication for increasing soybean seed weight in molecular design breeding. Cloning this locus in future may provide new insights into the genetic mechanisms underlying soybean seed size traits.

Variation in seed and seedling traits and their relations to geo-climatic factors among populations in Yunnan Pine(Pinus yunnanensis)

Differences in seed germination and seedling growth among populations of Pinus yunnanensis Franch. were quantified to provide information for use in the regeneration, reforestation, and improvement of this species. Seed germination and seedling growth traits at the nursery stage were investigated at the population level. We evaluated relationships between pairs of measured traits as well as the association between measured traits and geo- climatic variables including longitude, latitude, elevation, temperature and precipitation. Seedling growth showed significant differences although a low magnitude of variation was found in the seed germination traits among the populations. Some populations with strong and stable growth （e.g., the Baoshan population） may be selected as superior seed sources for regeneration and reforestation. Germination percentage was negatively correlated with elevation, and positively with precipitation. In addition, germination index, vigor index and seedling height were positively correlated with precipitation. Seed germination index, cotyledon number and length, and seedling diameter were significantly and positively correlated with seed weight. Strong correlations between pairs of traits will be helpful in early evaluations for the selection of certain prominent traits.

Lychee seed saponins improve cognitive function and ameliorate hippocampal neuronal injury in Alzheimer disease rat model induced by Aβ_(25-35) through AKT/GSK3β pathway

OBJECTIVE Lychee seed,a famous traditional Chinese medicine,recently were reported to improve the learning and memory abilities in mice.However,it is still unclear whether lychee seed saponins(LSS)can improve the cognitive function and associated mechanisms.METHODS In present studies,we established the Alzheimer disease(AD)model by injecting Aβ25-35 into the lateral ventricle of rats.Then the spatial learning and memory abilities of LSS-treated rats were evaluated with the Morris water maze,meanwhile the protein expressions of AKT,GSK3β and Tau in the hippocampal neuron were analyzed by immunohistochemistry and Western blotting.RESULTS The results showed LSS can improve the cognitive functions of AD rats through shortening the escape latency,increasing the number across the platform,platform quadrant dwell time and the percentage of the total distance run platform quadrant.The protein expression of AKT was significantly up-regulated and that of GSK3β and Tau were decreased remarkably in the hippocampal CA1 area.CONCLUSION Our study is the first to show that LSS significantly improve the cognitive function and prevent hippocampal neuronal injury of the rats with AD by activation of the PI3K/AKT/GSK3βsignaling pathway,suggesting LSS may be developed into the nutrient supplement for the treatment of AD.

Relationship between miR-7-5p expression and ^(125)I seed implantation efficacy in pancreatic cancer and functional analysis of target genes

Objective The aim of this study was to investigate the relationship between miR-7-5p expression and intertissue-^(125)I irradiation sensitivity in pancreatic cancer tissues and to analyze the function of target genes.Methods Thirty-seven patients with unresectable pancreatic ductal adenocarcinoma(PDAC)treated with radioactive ^(125)I seed implantation were enrolled.RT-PCR was used to detect the expression level of miR-7-5p in cancer tissues and analyze the relationship between miR-7-5p expression and ^(125)I radiation sensitivity.Bioinformatic software and online tools were used to predict the miR-7-5p target genes and analyze their functional annotation and pathway enrichment.Results Radioactive ^(125)I seed implantation was followed up for 2 months.The objective response rate of the miR-7-5p high expression group was 65.0%(13/20),whereas the objective response rate of the miR-7-5p low expression group was 5.88%(1/17),and the difference between the two groups was statistically significant(χ^(2)=13.654,P<0.001).A total of 187 target genes were predicted using three databases.GO functional annotation showed that target genes were mainly involved in cellular response to insulin stimulus,regulation of gene expression by genetic imprinting,cytosol,peptidyl-serine phosphorylation,bHLH transcription factor binding,cargo loading into vesicles,cellular response to epinephrine stimulus,and nucleoplasm.KEGG pathway enrichment analysis showed that target genes were mainly involved in the ErbB signaling pathway,HIF-1 signaling pathway,axon guidance,longevity regulatory pathway,endocrine resistance,glioma,choline metabolism in cancer,and EGFR tyrosine kinase inhibitor drug resistance.Molecular complex detection analysis by Cytoscape revealed that PIGH,RAF1,EGFR,NXT2,PIK3CD,PIK3R3,ERBB4,TRMT13,and C5orf22 were the key modules of miR-7-5p target gene clustering.Conclusion The expression of miR-7-5p in pancreatic cancer tissues positively correlated with the radiosensitivity of ^(125)I seeds.Via targeted gene regulation,miR-7-5p acts on the network of multiple signaling pathways in PDAC and participates in its occurrence and development.Thus,miR-7-5p may become a predictive index of ^(125)I seed implantation therapy sensitivity in PDAC patients.

Innovative use of intact seeds of Mucuna monosperma Wight for improved yield of L-DOPA

The drug L-DOPA has been widely used against Parkinson's disease and is extracted from plants.Due to the increasing demand of this drug,new plant sources need to be discovered in addition to the existing sources.The paper embodies results on Mucuna monosperma,which can be a promising candidate for L-DOPA.The seed powder of this plant contains 5.48%of(dry weight)the drug and when the seeds were soaked in distilled water,content was increased to 6.58%.Different elicitors when added,enhanced the drug level in seed up to 11.8%.The possible rationale behind this increase was confirmed by increase in tyrosinase activity in the seeds.Presence of L-DOPA was confirmed using various analytical techniques as HPLC,HPTLC and NMR.The work demonstrates a potential candidate plant as a source for L-DOPA when a novel method was adopted as described here.

Quantitative Trait Loci Underlying Seed Sugars Content in “MD96-5722” by “Spencer” Recombinant Inbred Line Population of Soybean

Sucrose, raffinose, and stachyose are important soluble sugars in soybean [Glycine max (L.) Merr.] seeds. Seed sucrose is a desirable trait for taste and flavor. Raffinose and stachyose are undesirable in diets of monogastric animals, acting as anti-nutritional factors that cause flatulence and abdominal discomfort. Therefore, reducing raffinose and stachyose biosynthesis is considered as a key quality trait goal in soy food and feed industries. The objective of this study was to identify genomic regions containing quantitative trait loci (QTL) controlling sucrose, raffinose, and stachyose in a set of 92 F5:7 recombinant inbred lines (RILs) derived from a cross between the lines “MD96-5722” and “Spencer” by using 5376 Single Nucleotide Polymorphism (SNP) markers from the Illumina Infinium SoySNP6K BeadChip array. Fourteen significant QTL were identified and mapped on eight different linkage groups (LGs) and chromosomes (Chr). Three QTL for seed sucrose content were identified on LGs N (Chr3), K (Chr9), and E (Chr15). Seven QTL were identified for raffinose content on LGs D1a (Chr1), N (Chr3), C2 (Chr6), K (Chr9), B2 (Chr14), and J (Chr16). Four QTL for stachyose content were identified on LG D1a (Chr1), C2 (Chr6), H (Chr12), and B2 (Chr14). Selection for beneficial alleles of these QTLs could facilitate breeding strategies to develop soybean lines with higher concentrations of sucrose and lower levels of raffinose and stachyose.

PEG-6000引发对白羊草种子发芽的影响

为了探讨PEG-6000溶液引发对白羊草种子萌发的影响,采用水势0、-0.2、-0.4和-0.6Mpa的聚乙二醇溶液对白羊草种子进行引发0、3、6和9 h处理,测定白羊草种子的发芽率和发芽势,并计算其发芽指数、活力指数、平均发芽天数和发芽速率系数。试验结果表明:(1)水势为-0.6 Mpa的PEG-6000溶液引发可以有效的提高白羊草种子的发芽速度系数至0.387,显著高于其他水势处理(P<0.05),并且在此水势处理下,引发3 h的发芽速度系数显著高于其他处理(P<0.05);(2)不同引发时间均可以显著提高白羊草种子的发芽能力(P<0.05),引发6 h比引发3 h和9 h的白羊草种子发芽能力高,但差异不显著。

^(60)Co-γ对不同遗传背景甘蓝型黄籽油菜种子发芽力的影响

以11个不同遗传背景的甘蓝型黄籽油菜品系为材料，分别选出同一遗传品系的黄、黑籽给予120GY、1600GY、2000GY、240GY剂量的60Co－γ射线辐照，测定其发芽势和发芽率，并进行方差分析。结果表明，不同遗传背景的品系间、颜色间、剂量间的发芽力存在着极显著差异。少数品系的黄籽耐辐射能力优于黑籽，通过筛选，可望从中选育出抗逆性强的优良黄籽品系。

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。

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.

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。

单籽鉴定法筛选辣椒黄绿苗胞质雄性不育系YBM-A6恢复系

为了简化辣椒黄绿苗胞质雄性不育系恢复系的选育方法,春季以辣椒黄绿苗胞质雄性不育系YBM-A6作为母本与210个自交系进行杂交,当年秋季将得到的F1种子随机取2粒饱满的种子进行单籽法筛选,至开出3朵花后开始鉴定其育性;第2年春季进行大田播种鉴定,鉴定内容与单籽鉴定法相同。结果表明:2种鉴定方法鉴定结果相近,单籽鉴定法及大田鉴定筛选出的恢复系占总试验材料的百分比分别为24.8%和24.3%,单籽鉴定法较大田鉴定法能够大量节省鉴定用地,节省近9个月鉴定时间,并且节省鉴定费用。

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。

GC-MS分析黄褐籽甘蓝型油菜种子发育过程中脂肪酸含量变化

利用气相色谱质谱(GC-MS)联用分析技术,对细胞工程创建的甘蓝型油菜黄籽株系W82和其褐籽亲本‘扬油6号'种子发育过程中脂肪酸的含量进行了分析。发现在黄/褐籽油菜种子发育早期,随着种子的发育脂肪酸逐渐积累,在授粉后第5周(WAF5,week after fertilization)时含量开始下降,在WAF6含量又逐渐上升。黄/褐籽甘蓝型油菜种子在发育早期主要积累碳链较短的脂肪酸,在后期主要积累碳链较长的不饱和脂肪酸。黄籽株系W82种子中的脂肪酸含量要高于褐籽亲本‘扬油6号'。