套作大豆苗期茎秆纤维素合成代谢与抗倒性的关系

为从茎秆强度的角度研究套作大豆苗期对荫蔽胁迫的响应及耐荫抗倒机制,采用耐荫性不同的3个大豆材料,在玉米大豆套作和单作两种种植模式下,对茎秆的纤维素、可溶性糖、蔗糖、淀粉含量及蔗糖代谢中关键酶活性以及茎秆抗折力、抗倒伏指数等进行测定,研究它们与套作大豆苗期倒伏的关系.套作大豆苗期倒伏严重,茎秆抗折力、抗倒伏指数、纤维素、可溶性糖、蔗糖、淀粉含量和相关酶活性均显著低于单作.不同大豆材料受套作荫蔽影响程度不同,强耐荫性大豆南豆12茎秆抗折力降低幅度最小,在套作环境下其茎秆抗折力、抗倒伏指数大,纤维素、可溶性糖、蔗糖、淀粉含量高,酶活性强.相关分析表明： 套作大豆苗期茎秆糖含量均与抗折力呈极显著正相关,与倒伏率呈极显著负相关;蔗糖含量与蔗糖磷酸合酶(SPS)、蔗糖合酶(SS)、中性转化酶(NI)活性呈极显著正相关,与酸性转化酶(AI)活性相关性不显著;纤维素含量与SPS、SS呈极显著正相关,与NI呈显著正相关,与AI相关性不显著.套作环境下,强耐荫性大豆苗期茎秆中较高的SPS、SS活性是其维持高蔗糖和纤维素含量的酶学基础,而高纤维素含量有利于提高茎秆强度,进而增强其抗倒伏能力.本研究应用玉米大豆套作种植系统,从苗期抗倒角度,探明了光环境对不同基因型大豆茎秆纤维素代谢的影响机制,为下一步筛选耐荫抗倒大豆品种提供了理论依据.     

甘蓝型油菜扩展蛋白家族的全基因组鉴定及其对缺硼胁迫响应的差异分析

以甘蓝型油菜(Brassica napus L.)硼高效品种‘青油10号’和硼低效品种‘Westar 10’为研究对象,采用生物信息学分析、转录组测序和实时荧光定量PCR技术,鉴定其基因组中扩展蛋白的家族成员,并对该基因家族响应缺硼胁迫的表达差异进行分析。结果显示,甘蓝型油菜基因组中包含109个扩展蛋白,可分为4个亚家族,包括:79个扩展蛋白A(BnaEXPAs)、21个扩展蛋白B(BnaEXPBs)、5个类扩展蛋白A(BnaEXLAs)和4个类扩展蛋白B(BnaEXLBs)。同一亚家族中的扩展蛋白具有相对保守的基因结构和蛋白质基序组成。这些扩展蛋白基因分布在19条染色体上,其中10个位于硼高效QTL区间内。转录组测序分析结果表明,缺硼胁迫时‘青油10号’的根、幼叶和老叶中分别有40、18和30个扩展蛋白基因显著上调或下调表达;而‘Westar10’中分别有27、24和41个扩展蛋白基因显著上调或下调表达。其中‘青油10号’根中的BnaC04.EXPA6a,幼叶中的BnaA09.EXPA5以及老叶中的BnaA09.EXPA16、BnaC04.EXPA3、BnaCnn.EXPA5b和BnaA03.EXPA8基因的表达水平均显著高于‘Westar10’。研究结果说明甘蓝型油菜基因组中扩展蛋白基因家族数量庞大,其中高、低效品种间和不同硼水平中差异表达的扩展蛋白可能在甘蓝型油菜低硼适应性中发挥重要作用。     

水稻脆秆突变体bc-s1的表型分析和基因定位

茎秆机械强度影响植株抗倒伏能力, 是备受关注的重要农艺性状之一。与野生型相比, 水稻(Oryza sativa)脆秆隐性突变体bc-s1茎秆抗折力和抗张力分别降低31.1%和67.2%, 茎秆纤维素和木质素含量分别降低24.97%和增高38.82%。细胞学分析显示, bc-s1茎秆厚壁细胞发生不规则变化, 次生壁增厚受阻。通过图位克隆和测序分析, 初步确定bc-s1突变体中纤维素合成酶催化亚基Os09g25490/OsCesA9基因第1外显子的第28个碱基G突变为A。该等位突变体的获得为进一步揭示OsCesA9调控细胞壁建成的生物学功能提供了新的研究材料。     

种植密度与喷施多效唑对冬小麦抗倒伏能力和产量的影响

于2008-2010年2个生长季在山东农业大学试验农场,以烟农21和藁城8901为材料,研究了不同种植密度下喷施多效唑对冬小麦基部茎秆形态特征、抗折力和抗倒指数及产量的影响.结果表明：藁城8901的抗倒伏能力高于烟农21,但籽粒产量低.与低密度（基本苗为180×10⁴株·hm^-2）条件相比,同一品种高密度（基本苗为240×10⁴株·hm^-2）使茎秆抗折力和抗倒指数下降,尤以烟农21表现更为明显.喷施多效唑可明显降低株高和基部节间长度,显著提高茎秆抗折力和抗倒伏指数,增强其抗倒伏能力,提高单位面积穗数和产量.相关分析表明,茎秆抗倒伏指数与基部第2节间长度、基部(1+2)节间长度占节间总长的比例和表观倒伏率均呈显著负相关.因此,采用合理的种植密度并结合喷施多效唑,能够提高小麦的抗倒伏能力和产量,可作为半湿润地区小麦高产栽培的重要技术措施.     

不同养分和水分管理模式对水稻抗倒伏能力的影响

大田试验下,通过对水稻茎秆基部物理性状、形态特征和硅、钾含量的测定和比较,系统研究了不同养分和水分管理模式对水稻茎秆抗倒伏能力的影响．结果表明,有机无机肥配施,特别是秸秆与化肥配施(CS)养分模式可明显提高水稻植侏茎秆粗度、茎壁厚度和茎重,从而有效提高了基部茎秆的抗折力(RS)和明显降低了水稻的倒伏指数(LI),秸秆与化肥配施(CS)养分模式对水稻抗倒伏能力的效果在干湿交替(AWD)和控水模式(DRA)下表现更为明显。有机无机肥配施,特别是秸秆与化肥配施(CS)的养分模式在干湿交替(AWD)和控水模式(DRA)下更有助于提高水稻茎秆的硅、钾含量、相关分析表明,水稻基部茎秆茎壁厚度、茎重和抗折力与茎秆硅、钾含量存在显著(P<0．05)或极显著正相关(P<0．01)。     

甘蓝型油菜每角果粒数全基因组关联分析

为挖掘甘蓝型油菜每角果粒数显著关联单核苷酸多态性（SNP）位点及相关候选基因。本研究以300份甘蓝型油菜自交系为试验材料,对甘蓝油菜每角果粒数进行一年两地表型考察,并结合该群体前期开发的201 817个SNPs标记,采用一般线性模型（GLM）和混合线性模型（MLM）进行全基因组关联分析（GWAS）,此外,对性状显著关联SNP位点两侧100 kb区域内相关候选基因进行功能预测。300份甘蓝型油菜每角果粒数在两地均表现出广泛的表型变异,筛选出2份每角果粒数较多的油菜种质资源。基于GLM模型检测到39个与油菜每角果粒数显著关联SNPs,采用MLM分析发现,两地共检测到的3个每角果粒数显著关联SNPs位点均在GLM检测到。8个位点附近找到CIK,ERF022和EDE1等19个拟南芥已报道角果籽粒发育相关的同源基因。研究结果有助于解析甘蓝型油菜每角果粒数的遗传基础,为研究每角果粒数的调控机制、指导每角果粒数的遗传改良奠定基础。     

水稻不同生育期茎秆生化成分的变化及其与抗倒伏能力的关系

以抗倒伏品种‘南粳44’、‘武运粳7号’与不抗倒伏品系‘宁7412’为试验材料,通过对水稻不同生育期茎秆硅、钾、钙、镁含量及可溶性糖含量的测定,结合氮钾肥配比试验,研究了水稻不同生育期茎秆硅、钾、钙、镁含量及可溶性糖含量的变化及其与茎秆抗倒伏能力的关系。结果表明：水稻茎秆的硅、钙、镁含量及可溶性糖含量随生育进程呈上升趋势,而茎秆的钾含量呈现先降后升的趋势。在不同施肥水平条件下,水稻茎秆的硅、钾、钙、镁含量及可溶性糖含量存在一定的差异。抗倒伏品种‘南粳44’和‘武运粳7号’茎秆基部抗折力较强,施肥对水稻茎秆基部抗折力有一定的影响。水稻生殖生育期茎秆的硅含量、可溶性糖含量与茎秆基部抗折力呈极显著正相关（P〈0.01）,与水稻抗倒伏能力的强弱有一定的关系。     

RFLP和AFLP分析白菜型油菜和甘蓝型油菜遗传多样性及其在油菜改良中的应用价值

采用RFLP和AFLP标记对来自中国和欧美的7份甘蓝型油菜和22份白菜型油菜进行了遗传多样性分析。在这29份材料中,166个酶-探针组合和2对AFLP引物共检测到1477个RFLP标记和183个AFLP标记。RFLP数据显示以拟南芥EST克隆作探针比用油菜基因组克隆做探针能检测到更多的多态性位点,且采用EcoR Ⅰ或BamH Ⅰ酶切比HindⅢ酶切多态性好,白菜型油菜和甘蓝型油菜中基因的拷贝数平均都为3个左右。UPGMA聚类分析表明中国白菜型油菜的遗传多样性比甘蓝型油菜和欧美白菜型油菜丰富,欧美甘蓝型油菜与欧美白菜型油菜聚为一类,而与中国甘蓝型油菜差异更大。中国白菜型油菜丰富的遗传多样性为中国甘蓝型油菜的改良提供了宝贵的资源,揭示了利用白菜型油菜A基因组和甘蓝型油菜A基因组间亚基因组杂种优势的可能性。     

硅钾肥配施对水稻茎秆理化性状及抗倒伏能力的影响

该研究以易倒伏品种‘B优827’为试验材料,通过分析硅钾肥配合施用对水稻茎秆的弯曲力矩、抗折力以及倒伏指数的影响,探索通过肥料调控提高水稻抗倒伏能力的方法与技术措施。结果显示:(1)硅钾肥配施能显著增加水稻基部节间茎秆直径和壁厚,缩短节间长度,并在低硅中钾处理(T2,硅肥300kg/hm2、钾肥400kg/hm2)下基部各节间长度缩短幅度最大(12.44%~20.80%),直径和壁厚增加幅度分别为2.82%~5.76%、22.95%~28.57%。(2)硅钾肥配施处理后,水稻基部各节间纤维素、木质素及灰分含量显著变化,并以低硅中钾处理(T2)差异较显著,且基部节间木质素含量分别比对照显著提高14.55%、8.67%、7.73%。(3)硅钾肥配施处理能显著增加水稻基部节间抗折力,降低倒伏指数,同时仍以低硅中钾处理(T2)的抗折力最大、倒伏指数最小,对提高‘B优827’植株抗倒伏能力效果最好。研究表明,合理的硅钾肥配比(硅肥300kg/hm2、钾肥400kg/hm2)能显著增加易倒伏水稻品种‘B优827’的茎秆壁厚和直径,缩短基部节间长度,提高抗折力,同时还能增加其基部节间木质化程度,最终增强植株抗倒伏能力;硅钾肥配合施用可通过改善茎秆物理化学特性有效增强易倒伏水稻植株的抗倒伏能力。     

油菜矮秆突变WRKY转录因子cDNA克隆及表达分析

以甘蓝型油菜为材料,利用已建立的抑制性消减文库(SSH),采用RACE技术克隆到1个植物WRKY转录因子相关基因,命名为BnD11,其cDNA全长1034 bp,含有810 bp的完整开放阅读框,编码269个氨基酸。该基因编码的氨基酸序列与拟南芥WRKY40氨基酸序列相似性为79%,与拟南芥中编码WRKY-DNA结合蛋白40基因的氨基酸序列相似性达78%,与其它多种植物的WRKY转录因子的氨基酸序列也有较高的相似性。半定量RT-PCR对BnD11进行组织特异性表达分析显示:在正常生长条件下,BnD11在野生型和矮秆油菜的各个组织中均有表达,但在矮秆突变的根、茎、茎尖的相对表达量明显高于野生型。研究表明,BnD11功能区段具有很高的保守性,可能参与了油菜的茎秆发育。     

Sequence,expression divergence,and complementation of homologous <Emphasis Type="Italic">ALCATRAZ</Emphasis> loci in <Emphasis Type="Italic">Brassica napus</Emphasis>

The genomic era provides new perspectives in understanding polyploidy evolution, mostly on the genome-wide scale. In this paper, we show the sequence and expression divergence between the homologous ALCATRAZ (ALC) loci in Brassica napus, responsible for silique dehiscence. We cloned two homologous ALC loci, namely BnaC.ALC.a and BnaA.ALC.a in B. napus. Driven by the 35S promoter, both the loci complemented to the alc mutation of Arabidopsis thaliana, yet only the expression of BnaC.ALC.a was detectable in the siliques of B. napus. Sequence alignment indicated that BnaC.ALC.a and BolC.ALC.a, or BnaA.ALC.a and BraA.ALC.a, possess a high level of similarity. The understanding of the sequence and expression divergence among homologous loci of a gene is of due importance for an effective gene manipulation and TILLING (or ECOTILLING) analysis for the allelic DNA variation at a given locus. S. Hua and I. H. Shamsi contributed equally to this work.     

Substrates of the chloroplast small heat shock proteins 22E/F point to thermolability as a regulative switch for heat acclimation in <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis>

We previously described a Brassica napus chlorophyll-deficient mutant (ygl) with yellow-green seedling leaves and mapped the related gene, BnaC.YGL, to a 0.35 cM region. However, the molecular mechanisms involved in this chlorophyll defect are still unknown. In this study, the BnaC07.HO1 gene (equivalent to BnaC.YGL) was isolated by the candidate gene approach, and its function was confirmed by genetic complementation. Comparative sequencing analysis suggested that BnaC07.HO1 was lost in the mutant, while a long noncoding-RNA was inserted into the promoter of the homologous gene BnaA07.HO1. This insert was widely present in B. napus cultivars and down-regulated BnaA07.HO1 expression. BnaC07.HO1 was highly expressed in the seedling leaves and encoded heme oxygenase 1, which was localized in the chloroplast. Biochemical analysis showed that BnaC07.HO1 can catalyze heme conversion to form biliverdin IXα. RNA-seq analysis revealed that the loss of BnaC07.HO1 impaired tetrapyrrole metabolism, especially chlorophyll biosynthesis. According, the levels of chlorophyll intermediates were reduced in the ygl mutant. In addition, gene expression in multiple pathways was affected in ygl. These findings provide molecular evidences for the basis of the yellow-green leaf phenotype and further insights into the crucial role of HO1 in B. napus.     

Genome-wide association study dissects the genetic control of plant height and branch number in response to low-phosphorus stress in Brassica napus

Background and AimsOilseed rape (Brassica napus) is one of the most important oil crops worldwide. Phosphorus (P) deficiency severely decreases the plant height and branch number of B. napus. However, the genetic bases controlling plant height and branch number in B. napus under P deficiency remain largely unknown. This study aims to mine candidate genes for plant height and branch number by genome-wide association study (GWAS) and determine low-P-tolerance haplotypes.MethodsAn association panel of B. napus was grown in the field with a low P supply (P, 0 kg ha⁻¹) and a sufficient P supply (P, 40 kg ha⁻¹) across 2 years and plant height and branch number were investigated. More than five million single-nucleotide polymorphisms (SNPs) were used to conduct GWAS of plant height and branch number at two contrasting P supplies.Key ResultsA total of 2127 SNPs were strongly associated (P < 6·25 × 10⁻⁰⁷) with plant height and branch number at two P supplies. There was significant correlation between phenotypic variation and the number of favourable alleles of associated loci on chromosomes A10 (chrA10_821671) and C08 (chrC08_27999846), which will contribute to breeding improvement by aggregating these SNPs. BnaA10g09290D and BnaC08g26640D were identified to be associated with chrA10_821671 and chrC08_27999846, respectively. Candidate gene association analysis and haplotype analysis showed that the inbred lines carrying ATT at BnaA10g09290Hap1 and AAT at BnaC08g26640Hap1 had greater plant height than lines carrying other haplotype alleles at low P supply.ConclusionOur results demonstrate the power of GWAS in identifying genes of interest in B. napus and provided insights into the genetic basis of plant height and branch number at low P supply in B. napus. Candidate genes and favourable haplotypes may facilitate marker-based breeding efforts aimed at improving P use efficiency in B. napus.     

<Emphasis Type="Italic">LMI1</Emphasis>-like genes involved in leaf margin development of <Emphasis Type="Italic">Brassica napus</Emphasis>

In rapeseed (Brassica napus L.), leaf margins are variable and can be entire, serrate, or lobed. In our previous study, the lobed-leaf gene (LOBED-LEAF 1, BnLL1) was mapped to a 32.1 kb section of B. napus A10. Two LMI1-like genes, BnaA10g26320D and BnaA10g26330D, were considered the potential genes that controlled the lobed-leaf trait in rapeseed. In the present study, these two genes and another homologous gene (BnaC04g00850D) were transformed into Arabidopsis thaliana (L.) Heynh. plants to identify their functions. All three LMI1-like genes of B. napus produced serrate leaf margins. The expression analysis indicated that the expression level of BnaA10g26320D determined the difference between lobed- and entire-leaved lines in rapeseed. Therefore, it is likely that BnaA10g26320D corresponds to BnLL1.     

Determination of fatty acid composition in seed oil of rapeseed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.) by mutated alleles of the FAD3 desaturase genes

One of the goals in oilseed rape programs is to develop genotypes producing oil with low linolenic acid content (C18:3, ≤3%). Low linolenic mutant lines of canola rapeseed were obtained via chemical mutagenesis at the Plant Breeding and Acclimatization Institute – NRI, in Poznan, Poland, and allele-specific SNP markers were designed for monitoring of two statistically important single nucleotide polymorphisms detected by SNaPshot analysis in two FAD3 desaturase genes, BnaA.FAD3 and BnaC.FAD3, respectively. Strong negative correlation between the presence of mutant alleles of the genes and linolenic acid content was revealed by analysis of variance. In this paper we present detailed characteristics of the markers by estimation of the additive and dominance effects of the FAD3 genes with respect to particular fatty acid content in seed oil, as well as by calculation of the phenotypic variation of seed oil fatty acid composition accounted by particular allele-specific marker. The obtained percentage of variation in fatty acid composition was considerable only for linolenic acid content and equaled 35.6% for BnaA.FAD3 and 39.3% for BnaC.FAD3, whereas the total percentage of variation in linolenic acid content was 53.2% when accounted for mutations in both genes simultaneously. Our results revealed high specificity of the markers for effective monitoring of the wild-type and mutated alleles of the Brassica napus FAD3 desaturase genes in the low linolenic mutant recombinants in breeding programs.