Genome-Wide Differential DNA Methylation and miRNA Expression Profiling Reveals Epigenetic Regulatory Mechanisms Underlying Nitrogen-Limitation-Triggered Adaptation and Use Efficiency Enhancement in Allotetraploid Rapeseed.

Improving crop nitrogen (N) limitation adaptation (NLA) is a core approach to enhance N use efficiency (NUE) and reduce N fertilizer application. Rapeseed has a high demand for N nutrients for optimal plant growth and seed production, but it exhibits low NUE. Epigenetic modification, such as DNA methylation and modification from small RNAs, is key to plant adaptive responses to various stresses. However, epigenetic regulatory mechanisms underlying NLA and NUE remain elusive in allotetraploid B. napus. In this study, we identified overaccumulated carbohydrate, and improved primary and lateral roots in rapeseed plants under N limitation, which resulted in decreased plant nitrate concentrations, enhanced root-to-shoot N translocation, and increased NUE. Transcriptomics and RT-qPCR assays revealed that N limitation induced the expression of NRT1.1, NRT1.5, NRT1.7, NRT2.1/NAR2.1, and Gln1;1, and repressed the transcriptional levels of CLCa, NRT1.8, and NIA1. High-resolution whole genome bisulfite sequencing characterized 5094 differentially methylated genes involving ubiquitin-mediated proteolysis, N recycling, and phytohormone metabolism under N limitation. Hypermethylation/hypomethylation in promoter regions or gene bodies of some key N-metabolism genes might be involved in their transcriptional regulation by N limitation. Genome-wide miRNA sequencing identified 224 N limitation-responsive differentially expressed miRNAs regulating leaf development, amino acid metabolism, and plant hormone signal transduction. Furthermore, degradome sequencing and RT-qPCR assays revealed the miR827-NLA pathway regulating limited N-induced leaf senescence as well as the miR171-SCL6 and miR160-ARF17 pathways regulating root growth under N deficiency. Our study provides a comprehensive insight into the epigenetic regulatory mechanisms underlying rapeseed NLA, and it will be helpful for genetic engineering of NUE in crop species through epigenetic modification of some N metabolism-associated genes.

A multiomics approach reveals the pivotal role of subcellular reallocation in determining rapeseed resistance to cadmium toxicity.

Oilseed rape (Brassica napus) has great potential for phytoremediation of cadmium (Cd)-polluted soils due to its large plant biomass production and strong metal accumulation. Enhanced plant Cd resistance (PCR) is a crucial prerequisite for phytoremediation through hyper-accumulation of excess Cd. However, the complexity of the allotetraploid genome of rapeseed hinders our understanding of PCR. To explore rapeseed Cd-resistance mechanisms, we examined two genotypes, 'ZS11' (Cd-resistant) and 'W10' (Cd-sensitive), that exhibit contrasting PCR while having similar tissue Cd concentrations, and characterized their different fingerprints in terms of plant morphophysiology (electron microscopy), ion abundance (inductively coupled plasma mass spectrometry), DNA variation (whole-genome resequencing), transcriptomics (high-throughput mRNA sequencing), and metabolomics (ultra-high performance liquid chromatography-mass spectrometry). Fine isolation of cell components combined with ionomics revealed that more Cd accumulated in the shoot vacuoles and root pectins of the resistant genotype than in the sensitive one. Genome and transcriptome sequencing identified numerous DNA variants and differentially expressed genes involved in pectin modification, ion binding, and compartmentalization. Transcriptomics-assisted gene co-expression networks characterized BnaCn.ABCC3 and BnaA8.PME3 as the central members involved in the determination of rapeseed PCR. High-resolution metabolic profiles revealed greater accumulation of shoot Cd chelates, and stronger biosynthesis and higher demethylation of root pectins in the resistant genotype than in the sensitive one. Our comprehensive examination using a multiomics approach has greatly improved our understanding of the role of subcellular reallocation of Cd in the determination of PCR.

Balance between nitrogen use efficiency and cadmium tolerance in Brassica napus and Arabidopsis thaliana.

The transmembrane transport of NO3- and Cd2+ into plant cell vacuoles relies on the energy from their tonoplast proton pumps, V-ATPase and V-PPase. If the activity of these pumps is reduced, it results in less NO3- and Cd2+ being transported into the vacuoles, which contributes to better nitrogen use efficiency (NUE) and lower Cd2+ tolerance in plants. The physiological mechanisms that regulate the balance between NUE and Cd2+ tolerance remain unknown. In our study, two Brassica napus genotypes with differential NUEs, xiangyou 15 and 814, and Atclca-2 mutant and AtCAX4 over-expression line (AtCAX4-OE) of Arabidopsis thaliana, were used to investigate Cd2+ stress responses. We found that the Brassica napus genotype, with higher NUE, was more sensitive to Cd2+ stress. The AtCAX4-OE mutant, with higher Cd2+ vacuolar sequestration capacity (VSC), limited NO3- sequestration into root vacuoles and promoted NUE. Atclca-2 mutants, with decreased NO3- VSC, enhanced Cd2+ sequestration into root vacuoles and conferred greater Cd2+ tolerance than the WT. This may be due to the competition between Cd2+ andNO3- in the vacuoles for the energy provided by V-ATPase and V-PPase. Regulating the balance between Cd2+ and NO3- vacuolar accumulation by inhibiting the activity of CLCa transporter and increasing the activity of CAX4 transporter will simultaneously enhance both the NUE and Cd2+ tolerance of Brassica napus, essential for improving its Cd2+ phytoremediation potential.

Integrated physiologic, genomic and transcriptomic strategies involving the adaptation of allotetraploid rapeseed to nitrogen limitation.

BACKGROUND: Nitrogen (N) is a macronutrient that is essential for optimal plant growth and seed yield. Allotetraploid rapeseed (AnAnCnCn, 2n = 4x = 38) has a higher requirement for N fertilizers whereas exhibiting a lower N use efficiency (NUE) than cereal crops. N limitation adaptation (NLA) is pivotal for enhancing crop NUE and reducing N fertilizer use in yield production. Therefore, revealing the genetic and molecular mechanisms underlying NLA is urgent for the genetic improvement of NUE in rapeseed and other crop species with complex genomes. RESULTS: In this study, we integrated physiologic, genomic and transcriptomic analyses to comprehensively characterize the adaptive strategies of oilseed rape to N limitation stresses. Under N limitations, we detected accumulated anthocyanin, reduced nitrate (NO3-) and total N concentrations, and enhanced glutamine synthetase activity in the N-starved rapeseed plants. High-throughput transcriptomics revealed that the pathways associated with N metabolism and carbon fixation were highly over-represented. The expression of the genes that were involved in efficient N uptake, translocation, remobilization and assimilation was significantly altered. Genome-wide identification and molecular characterization of the microR827-NLA1-NRT1.7 regulatory circuit indicated the crucial role of the ubiquitin-mediated post-translational pathway in the regulation of rapeseed NLA. Transcriptional analysis of the module genes revealed their significant functional divergence in response to N limitations between allotetraploid rapeseed and the model Arabidopsis. Association analysis in a rapeseed panel comprising 102 genotypes revealed that BnaC5.NLA1 expression was closely correlated with the rapeseed low-N tolerance. CONCLUSIONS: We identified the physiologic and genome-wide transcriptional responses of oilseed rape to N limitation stresses, and characterized the global members of the BnamiR827-BnaNLA1s-BnaNRT1.7s regulatory circuit. The transcriptomics-assisted gene co-expression network analysis accelerates the rapid identification of central members within large gene families of plant species with complex genomes. These findings would enhance our comprehensive understanding of the physiologic responses, genomic adaptation and transcriptomic alterations of oilseed rape to N limitations and provide central gene resources for the genetic improvement of crop NLA and NUE.

Genome-scale characterization of the vacuole nitrate transporter Chloride Channel (CLC) genes and their transcriptional responses to diverse nutrient stresses in allotetraploid rapeseed.

The Chloride Channel (CLC) gene family is reported to be involved in vacuolar nitrate (NO3-) transport. Nitrate distribution to the cytoplasm is beneficial for enhancing NO3- assimilation and plays an important role in the regulation of nitrogen (N) use efficiency (NUE). In this study, genomic information, high-throughput transcriptional profiles, and gene co-expression analysis were integrated to identify the CLCs (BnaCLCs) in Brassica napus. The decreased NO3- concentration in the clca-2 mutant up-regulated the activities of nitrate reductase and glutamine synthetase, contributing to increase N assimilation and higher NUE in Arabidopsis thaliana. The genome-wide identification of 22BnaCLC genes experienced strong purifying selection. Segmental duplication was the major driving force in the expansion of the BnaCLC gene family. The most abundant cis-acting regulatory elements in the gene promoters, including DNA-binding One Zinc Finger, W-box, MYB, and GATA-box, might be involved in the transcriptional regulation of BnaCLCs expression. High-throughput transcriptional profiles and quantitative real-time PCR results showed that BnaCLCs responded differentially to distinct NO3- regimes. Transcriptomics-assisted gene co-expression network analysis identified BnaA7.CLCa-3 as the core member of the BnaCLC family, and this gene might play a central role in vacuolar NO3- transport in crops. The BnaCLC members also showed distinct expression patterns under phosphate depletion and cadmium toxicity. Taken together, our results provide comprehensive insights into the vacuolar BnaCLCs and establish baseline information for future studies on BnaCLCs-mediated vacuolar NO3- storage and its effect on NUE.

Effective Genomic Selection in a Narrow-Genepool Crop with Low-Density Markers: Asian Rapeseed as an Example.

Genomic selection (GS) has revolutionized breeding for quantitative traits in plants, offering potential to optimize resource allocation in breeding programs and increase genetic gain per unit of time. Modern high-density single nucleotide polymorphism (SNP) arrays comprising up to several hundred thousand markers provide a user-friendly technology to characterize the genetic constitution of whole populations and for implementing GS in breeding programs. However, GS does not build upon detailed genotype profiling facilitated by maximum marker density. With extensive genome-wide linkage disequilibrium (LD) being a common characteristic of breeding pools, fewer representative markers from available high-density genotyping platforms could be sufficient to capture the association between a genomic region and a phenotypic trait. To examine the effects of reduced marker density on genomic prediction accuracy, we collected data on three traits across 2 yr in a panel of 203 homozygous Chinese semiwinter rapeseed ( L.) inbred lines, broadly encompassing allelic variability in the Asian genepool. We investigated two approaches to selecting subsets of markers: a trait-dependent strategy based on genome-wide association study (GWAS) significance thresholds and a trait-independent method to detect representative tag SNPs. Prediction accuracies were evaluated using cross-validation with ridge-regression best linear unbiased predictions (rrBLUP). With semiwinter rapeseed as a model species, we demonstrate that low-density marker sets comprising a few hundred to a few thousand markers enable high prediction accuracies in breeding populations with strong LD comparable to those achieved with high-density arrays. Our results are valuable for facilitating routine application of cost-efficient GS in breeding programs.

Genomics-Assisted Identification and Characterization of the Genetic Variants Underlying Differential Nitrogen Use Efficiencies in Allotetraploid Rapeseed Genotypes.

Nitrogen (N) is a non-mineral macronutrient essential for plant growth and development. Oilseed rape (AnAnCnCn, 2n = 4x = 38) has a high requirement for N nutrients whereas showing the lowest N use efficiency (NUE) among crops. The mechanisms underlying NUE regulation in Brassica napus remain unclear because of genome complexity. In this study, we performed high-depth and -coverage whole-genome re-sequencing (WGS) of an N-efficient (higher NUE) genotype "XY15" and an N-inefficient (lower NUE) genotype "814" of rapeseed. More than 687 million 150-bp paired-end reads were generated, which provided about 93% coverage and 50× depth of the rapeseed genome. Applying stringent parameters, we identified a total of 1,449,157 single-nucleotide polymorphisms (SNPs), 335,228 InDels, 175,602 structure variations (SVs) and 86,280 copy number variations (CNVs) between the N-efficient and -inefficient genotypes. The largest proportion of various DNA polymorphisms occurred in the inter-genic regions. Unlike CNVs, the SNP/InDel and SV polymorphisms showed variation bias of the An and Cn subgenomes, respectively. Gene ontology analysis showed the genetic variants were mapped onto the genes involving N compound transport and ATPase complex metabolism, but not including N assimilation-related genes. On basis of identification of N-starvation responsive genes through high-throughput expression profiling, we also mapped these variants onto some key NUE-regulating genes, and validated their significantly differential expression between the N-efficient and -inefficient genotypes through qRT-PCR assays. Our data provide genome-wide high resolution DNA variants underlying NUE divergence in allotetraploid rapeseed genotypes, which would expedite the effective identification and functional validation of key NUE-regulating genes through genomics-assisted improvement of crop nutrient efficiency.

Effects of a controlled-release fertilizer on yield, nutrient uptake, and fertilizer usage efficiency in early ripening rapeseed (Brassica napus L.).

BACKGROUND: Nitrogen (N), phosphorous (P), and potassium (K) are critical nutrient elements necessary for crop plant growth and development. However, excessive inputs will lead to inefficient usage and cause excessive nutrient losses in the field environment, and also adversely affect the soil, water and air quality, human health, and biodiversity. METHODS: Field experiments were conducted to study the effects of controlled-release fertilizer (CRF) on seed yield, plant growth, nutrient uptake, and fertilizer usage efficiency for early ripening rapeseed (Xiangzayou 1613) in the red-yellow soil of southern China during 2011-2013. It was grown using a soluble fertilizer (SF) and the same amounts of CRF, such as SF1/CRF1 (3750 kg/hm2), SF2/CRF2 (3000 kg/hm2), SF3/CRF3 (2250 kg/hm2), SF4/CRF4 (1500 kg/hm2), SF5/CRF5 (750 kg/hm2), and also using no fertilizer (CK). RESULTS: CRF gave higher seed yields than SF in both seasons by 14.51%. CRF4 and SF3 in each group achieved maximum seed yield (2066.97 and 1844.50 kg/hm2, respectively), followed by CRF3 (1929.97 kg/hm2) and SF4 (1839.40 kg/hm2). There were no significant differences in seed yield among CK, SF1, and CRF1 (P>0.05). CRF4 had the highest profit (7126.4 CNY/hm2) and showed an increase of 12.37% in seed yield, and it decreased by 11.01% in unit fertilizer rate compared with SF4. The branch number, pod number, and dry matter weight compared with SF increased significantly under the fertilization of CRF (P<0.05). The pod number per plant was the major contributor to seed yield. On the other hand, the N, P, and K uptakes increased at first and then decreased with increasing the fertilizer rate at maturity, and the N, P, and K usage efficiency decreased with increasing the fertilizer rate. The N, P, and K uptakes and usage efficiencies of the CRF were significantly higher than those of SF (P<0.05). The N accumulation and N usage efficiency of CRF increased by an average of 13.66% and 9.74 percentage points, respectively, compared to SF. In conclusion, CRF significantly promoted the growth of rapeseed with using total N as the base fertilizer, by providing sufficient N in the later growth stages, and last by reducing the residual N in the soil and increasing the N accumulation and N usage efficiency.

Nitrogen Use Efficiency Is Mediated by Vacuolar Nitrate Sequestration Capacity in Roots of Brassica napus.

Enhancing nitrogen use efficiency (NUE) in crop plants is an important breeding target to reduce excessive use of chemical fertilizers, with substantial benefits to farmers and the environment. In Arabidopsis (Arabidopsis thaliana), allocation of more NO3 (-) to shoots was associated with higher NUE; however, the commonality of this process across plant species have not been sufficiently studied. Two Brassica napus genotypes were identified with high and low NUE. We found that activities of V-ATPase and V-PPase, the two tonoplast proton-pumps, were significantly lower in roots of the high-NUE genotype (Xiangyou15) than in the low-NUE genotype (814); and consequently, less vacuolar NO3 (-) was retained in roots of Xiangyou15. Moreover, NO3 (-) concentration in xylem sap, [(15)N] shoot:root (S:R) and [NO3 (-)] S:R ratios were significantly higher in Xiangyou15. BnNRT1.5 expression was higher in roots of Xiangyou15 compared with 814, while BnNRT1.8 expression was lower. In both B. napus treated with proton pump inhibitors or Arabidopsis mutants impaired in proton pump activity, vacuolar sequestration capacity (VSC) of NO3 (-) in roots substantially decreased. Expression of NRT1.5 was up-regulated, but NRT1.8 was down-regulated, driving greater NO3 (-) long-distance transport from roots to shoots. NUE in Arabidopsis mutants impaired in proton pumps was also significantly higher than in the wild type col-0. Taken together, these data suggest that decrease in VSC of NO3 (-) in roots will enhance transport to shoot and essentially contribute to higher NUE by promoting NO3 (-) allocation to aerial parts, likely through coordinated regulation of NRT1.5 and NRT1.8.

[Multifractal Analysis of Rapeseed Spectrum for Chlorophyll Diagnosis Modeling].

One of the most important topics in crop information science is how to make use of the crop's information for non-destructive nutrient diagnosis which can be solved with spectrum analysis. The canopy's spectrum feature is a key indicator to describe the nutritional status for the rapeseeds. The original spectrum is to be disturbed with external factors such as environment and climate; however, it is difficult to be directly used for rapeseed biomass diagnosis due to its huge fluctuation. However, the multifractal feature of the spectra remains stable relatively. In order to study the relationship between the canopy's spectrum of the rapeseed and its chlorophyll, based on the multifractal theory, a quantitative model of chlorophyll prediction and a qualitative model of planting pattern identification were proposed in this paper to study the high oleic acid rapeseed samples in 24 transplanting regions and 24 direct planting regions. At first, the generalized Hurst exponent and mass exponents together with other relevant multifractal parameters of the spectra were extracted with popular multifractal detrended fluctuation analysis (MF-DFA) in different six considered wavelength ranges. It shows that all of them possess representative multifractal nature. However, there are some differences of the multifractal characteristics between the two kinds of regions with different planting pattern in some bands. In addition, by correlation analysis and detection between the multifractal parameters of the spectra and the SPAD values in six considered ranges of bands, it demonstrates that there is some difference of the effective information content in the different ranges of bands. In the quantitative model of chlorophyll prediction, for each groups of samples in transplanting regions and direct planting regions and mixed together in each significant bands, a selected multifractal parameter was used to establish the univariate model for predicting the rapeseed leaf's SPAD values, respectively. The results of all the relative root mean square errors are small than 5%. Finally, the qualitative model was proposed to distinguish the samples by the two planting pattern. Youden index, as the identification accuracy was calculated for the six considered ranges of bands by the Fisher's linear discriminant analysis. The best Youden index is 0.902 5 and the corresponding band range is 350~1 350 nm. The significant work provides a theoretical and practical method for predicting rapeseed leaf's SPAD and also provides effective way to find the sensitive bands of the spectra for identification diagnosis.

[Effects of nitrogen application on yield and nitrogen use efficiency of rapeseed (Brassica napus)]. / èŠ±æœŸæ¸æ°´é€†å¢ƒä¸‹æ°®ç´ å¯¹æ²¹èœäº§é‡åŠæ°®è‚¥åˆ©ç”¨æ•ˆçŽ‡çš„å½±å“.

A greenhouse pot experiment was conducted to study the effects of nitrogen application on the yield and nitrogen use efficiency of rapeseed under post anthesis waterlogging condition. Two high nitrogen use efficiency rapeseed genotypes 'Monty' and 'Xiangyou 15' and two low nitrogen use efficiency rapeseed genotypes 'R210' and 'Bin270' were treated with 3 nitrogen levels (0.05, 0.2, 0.3 g N·kg-1soil) under waterlogging or normal water condition. The results showed that compared with the normal water condition, rapeseed pods per plant, 1000-seed mass, seed number per pod and seed yield decreased significantly under post anthesis waterlogging condition. Under the normal water condition, yield increased significantly along with the increment of nitrogen fertilizer, while under waterlogging condition the contribution of increment of nitrogen fertilizer was not significant. Compared with the low nitrogen use efficiency rapeseed genotypes, the high nitrogen use efficiency rapeseed genotypes stimulated seed filling under post anthesis waterlogging condition. Under the same water treatment, nitrogen use efficiency, nitrogen partial factor productivity, agronomic nitrogen use efficiency, nitrogen uptake efficiency and N harvest index of nitrogen fertilizer decreased significantly as the results of post anthesis waterlogging, nitrogen uptake and utilization capability of different rapeseed genotypes were affected significantly by waterlogging. Compared with the low nitrogen use efficiency rapeseed genotypes, the high nitrogen use efficiency rapeseed genotypes were more conducive to nitrogen translocation and redistribution of nitrogen to the pods under waterlogging condition, thus improving the seed production efficiency. Significant water and nitrogen interaction effects existed in rapeseed yield performance parameters, and the effects of water, nitrogen fertilizer and their interaction on rapeseed yield and yield performance parameters varied among diffe-rent genotypes.

[Effect of atmospheric CO2 concentration and nitrogen application level on absorption and transportation of nutrient elements in oilseed rape].

Effect of elevated atmospheric-CO2 (780 µmol . mol-1) on the absorption and transportation of secondary nutrient elements (calcium, magnesium, sulphur) and micronutrient elements (iron, manganese, zinc, molybdenum and boron) in oilseed rape at the stem elongation stage were studied by greenhouse simulated method. Compared with the ambient CO2 condition, the content of Zn in stem was increased and the contents of other nutrient elements were decreased under the elevated atmospheric-CO2 with no nitrogen (N) application; the contents of Ca, S, B and Zn were increased, and the contents of Mg, Mn, Mo and Fe were decreased under the elevated atmospheric CO2 with N application (0.2 g N . kg-1 soil); except the content of Mo in leaf was increased, the contents of other nutrient elements were decreased under the elevated atmospheric-CO2 with two levels of N application. Compared with the ambient CO2 condition, the amounts of Ca and S relative to the total amount of secondary nutrient elements in stem and the amounts of B and Zn relative to the total amount of micronutrient elements in stem were increased under the elevated-CO2 treatment with both levels of N application, and the corresponding values of Mg, Fe, Mn and Mo were decreased; no-N application treatment increased the proportion of Ca distributed into the leaves, and the proportion of Mg distributed into leaves was increased by the normal-N application level; the proportions of Mn, Zn and Mo distributed into the leaves were increased at both N application levels. Without N application, the elevation of atmospheric CO2 increased the transport coefficients of SFe, Mo and SS,B, but decreased the transport coefficients of SMg,Fe, SMg, Mn and SS,Fe, indicating the proportions of Mo, S transported into the upper part of plant tissues was higher than that of Fe, and the corresponding value of B was higher than that observed for S, the corresponding value of Mg was higher than that of Fe and Mn. Under normal-N application, the elevation of atmospheric CO2 increased the transport coefficients of SMg,Fe, SMg,Mn and SS,B, but decreased the transport coefficients of SCa, Mg, SFe,Mo and SS,Fe indicating the proportions of Fe, Mn and Ca transported into the upper part of plant tissues was higher than that of Mg; the corresponding value of B was higher than that observed for S, the corresponding value of Fe was higher than that of Mo, and the corresponding value of S was higher than that of Fe.

Characterization of the promoter and 5'-UTR intron of oleic acid desaturase (FAD2) gene in Brassica napus.

In the present study, we characterized the transcriptional regulatory region (KF038144) controlling the expression of a constitutive FAD2 in Brassica napus. There are multiple FAD2 gene copies in B. napus genome. The FAD2 gene characterized and analyzed in the study is located on chromosome A5 and was designated as BnFAD2A5-1. BnFAD2A5-1 harbors an intron (1,192 bp) within its 5'-untranslated region (5'-UTR). This intron demonstrated promoter activity. Deletion analysis of the BnFAD2A5-1 promoter and intron through the ß-glucuronidase (GUS) reporter system revealed that the -220 to -1 bp is the minimum promoter region, while -220 to -110 bp and +34 to +285 bp are two important regions conferring high-levels of transcription. BnFAD2 transcripts were induced by light, low temperature, and abscisic acid (ABA). These observations demonstrated that not only the promoter but also the intron are involved in controlling the expression of the BnFAD2A5-1 gene. The intron-mediated regulation is an essential aspect of the gene expression regulation.

[Sn-glycerol-3-phosphate acyltransferases (GPATs) in plants].

Sn-glycerol-3-phosphate acyltransferase (GPAT) catalyzes the acylation at sn-1 position of glycerol-3-phosphate to produce lysophosphatidic acid (LPA) in an acyl-CoA or acyl-ACP-dependent manner, which is the initial and rate-determining step of TAG biosynthetic pathway. Some GPATs have sn-2 transfer activity. Part members of the GPAT gene family have been cloned from different plant species. Based on their subcellular localizations, GPATs can be classified into three types, plastid GPATs, mitochondria GPATs and endoplasmic reticulum GPATs. GPATs exhibit diverse biochemical properties and are involved in synthesis of several lipids such as TAG, suberin, and cutin which play important roles in the growth and development of plants. This review summarized the current understanding of the chromosomal locus and gene structure of GPAT genes and the subcellular localization, sn-2 regiospecificity, substrates specialty, and functions of GPATs in plants.

[Molecular mapping and identification of quantitative trait loci for yield components in rapeseed (Brasscia napus L.)].

A F2 segregating population for genetic map construction and identification of QTL for seed yield in rapeseed (Brassica napus L.), was developed via crossing a conventional rapeseed line 04-1139 with a high yielding multiple silique rapeseed line 05-1054. A genetic map including 19 linkage groups was constructed with 200 SSR (Simple sequence repeat) and SRAP (Sequence-related amplified polymorphism) markers. This map covers a total length of 1 700.23 cM with an average distance between two adjacent makers of 8.50 cM. Using this map, QTL for the components of yield per plant, such as number of silique per plant (SNP), number of seeds per silique (SS) were analyzed. A total of 12 putative QTL for the traits were detected. Four of them were associated with SNP, which explained 35.64%, 12.96%, 28.71%, and 34.02% of the variation, respectively. Five QTL, which explained 8.41%, 7.87%, 24.37%, 8.57%, and 14.31% of the variation, were responsible for SS. Three QTL for 1 000-seed weight explained 1.81%-2.33% of the variation. The additional effects of the alleles for a trait may originate from both parents. The markers associated with the main QTL can be a good tool for marker associated selection and pyramiding breeding.

[Effects of chemical ripeners on chlorophyll content and antioxidant enzyme activities of rapeseed pod].

A field experiment was conducted to study the effects of ripeners Diguat and Roundup on the chlorophyll content, activities of catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD), cell membrane permeability, and malondialdehyde (MDA) content of rapeseed pods. Under effects of Diquat, the chlorophyll content decreased, while the activities of SOD, POD, and CAT, cell membrane permeability, and MDA content increased significantly, leading to the peroxidation of membrane lipid. These effects increased with increasing Diquat concentration. After treated with Roundup, the chlorophyll content had less change, activities of SOD, POD, and CAT increased slowly, and cell membrane permeability and MDA content had no obvious increase. With the increasing time of ripeners treatment, the activities of protective enzymes were inhibited to different degree, possibly due to the changes of molecular structure of antioxidant enzyme system under effects of the ripeners.

[Key enzymes in starch synthesis in plants].

Starch, the most common form of stored carbon in plants, is both the major food source for mankind and important raw material for many industries. It is composed of two types of alpha-1,4-linked glucan polymer: essentially unbranched amylose and regularly branched amylopectin, and synthesized in photosynthetic and non-photosynthetic organs. Starch is synthesized via four committed enzyme steps: ADP-Glc pyrophosphorylase, which synthesizes sugar nucleotide precursors; starch synthase, which extends the alpha-1,4-linked glucan chains using ADP-Glc; starch-branching enzymes, which introduce alpha-1,6 branch points to form amylopectin; and starch debranching enzymes, which hydrolyze alpha-1,6 branches in glucans. In this paper, recent advances in biochemical characterizations and gene engineering concerning these enzymes were reviewed, and the achievements in gene engineering involved in manipulation of starch amount and quality were also cited.