High-throughput root phenotyping screens identify genetic loci associated with root architectural traits in Brassica napus under contrasting phosphate availabilities

Background and Aims

Phosphate (Pi) deficiency in soils is a major limiting factor for crop growth worldwide. Plant growth under low Pi conditions correlates with root architectural traits and it may therefore be possible to select these traits for crop improvement. The aim of this study was to characterize root architectural traits, and to test quantitative trait loci (QTL) associated with these traits, under low Pi (LP) and high Pi (HP) availability in Brassica napus.

Methods

Root architectural traits were characterized in seedlings of a double haploid (DH) mapping population (n = 190) of B. napus [‘Tapidor’ × ‘Ningyou 7’ (TNDH)] using high-throughput phenotyping methods. Primary root length (PRL), lateral root length (LRL), lateral root number (LRN), lateral root density (LRD) and biomass traits were measured 12 d post-germination in agar at LP and HP.

Key Results

In general, root and biomass traits were highly correlated under LP and HP conditions. ‘Ningyou 7’ had greater LRL, LRN and LRD than ‘Tapidor’, at both LP and HP availability, but smaller PRL. A cluster of highly significant QTL for LRN, LRD and biomass traits at LP availability were identified on chromosome A03; QTL for PRL were identified on chromosomes A07 and C06.

Conclusions

High-throughput phenotyping of Brassica can be used to identify root architectural traits which correlate with shoot biomass. It is feasible that these traits could be used in crop improvement strategies. The identification of QTL linked to root traits under LP and HP conditions provides further insights on the genetic basis of plant tolerance to P deficiency, and these QTL warrant further dissection.     

Quantitative trait loci for root morphology in response to low phosphorus stress in Brassica napus

Phosphorus (P) deficiency in soils is a major limiting factor for crop growth worldwide. Changes in root morphology and architecture represent as an important mechanism of adaptation of plants to low P (LP) stress. To elucidate the genetic control of tolerance to P deficiency in Brassica napus, quantitative trait loci (QTL) for root morphology in response to LP were identified in three independent paper culture experiments, and dissected through QTL meta-analysis. In total, 62 significant QTL for total root length, root surface area, root volume, total dry weight, and plant P uptake under high and low P conditions were detected in the three experiments. Forty-five of these QTL were clustered within four linkage groups and were integrated into eight unique QTL by two rounds of QTL meta-analysis. Three of the unique QTL, uq.A1, uq.C3a and uq.C3b, were specific for LP condition. uq.C3a and uq.C3b were identified specifically for root traits and P uptake under LP stress, and may contribute to the adaptability of B. napus to P deficiency. Two functional markers, BnIPS2-C3 and BnGPT1-C3, which were developed from the genes AtIPS2 and AtGPT1 in Arabidopsis, were located in the confidence intervals of uq.C3a and uq.C3b, respectively. And AtGPT1 that corresponded to the interval of uq.C3b by in silico mapping was a possible candidate gene of uq.C3b. These results confirmed the importance of root traits for the adaptability of B. napus to LP and partially revealed the genetic basis of tolerance to P deficiency. These findings should be valuable for further study of the mechanism of P efficiency and the breeding of P-efficient cultivars by marker-assisted selection.     

Identification of quantitative trait loci associated with low boron stress that regulate root and shoot growth in Brassica napus seedlings

Brassica napus (Brassicaceae) is among the most important oil crops and a promising biofuel. In the tropics and subtropics, boron (B) deficiency is a major factor limiting Brassica yields. The effect of B on the regulation of root and shoot growth in a doubled haploid (DH) population was evaluated in experiments that utilized hydroponic culture. Strong genetic variability for traits of interest at normal and low B concentrations was demonstrated. Quantitative trait loci (QTL) were analyzed for seven plant growth parameters: increment of primary root length (IPRL), shoot dry weight (SDW), root dry weight (RDW), ratio of RDW to SDW (R/S), shoot B accumulation (SBA), root B accumulation (RBA), and ratio of RBA to SBA [(R/S)BA] in the population. Twenty-seven QTL were detected at normal B levels: four for IPRL, seven for SDW, three for RDW, two for R/S, six for SBA, two for RBA, and three for (R/S)BA. At low B, 18 QTL were detected: four for IPRL, three for SDW, two for RDW, two for R/S, five for SBA, one for RBA, and one for (R/S)BA. Three QTL for adaptability were detected: one A_IPRL and two A_SDW. No putative QTL was detected at both low and normal B. B-related genes were mapped in silico and their locations compared with the QTL identified. The present analyses show the profound and varied effects of B on B. napus and studies on QTL related to B efficiency will help to locate candidate genes and elucidate possible functions of B-efficiency-related QTL.     

Quantitative trait loci for seed yield and yield-related traits, and their responses to reduced phosphorus supply in Brassica napus

Background and Aims

One of the key targets of breeding programmes in rapeseed (Brassica napus) is to develop high-yield varieties. However, the lack of available phosphorus (P) in soils seriously limits rapeseed production. The aim of this study was to dissect the genetic control of seed yield and yield-related traits in B. napus grown with contrasting P supplies.

Methods

Two-year field trials were conducted at one site with normal and low P treatments using a population of 124 recombinant inbred lines derived from a cross between ‘B104-2’ and ‘Eyou Changjia’. Seed yield, seed weight, seed number, pod number, plant height, branch number and P efficiency coefficient (PEC) were investigated. Quantitative trait locus (QTL) analysis was performed by composite interval mapping.

Key Results

The phenotypic values of most of the tested traits were reduced under the low P conditions. In total, 74 putative QTLs were identified, contributing 7·3–25·4 % of the phenotypic variation. Of these QTLs, 16 (21·6 %) were detected in two seasons and in the mean value of two seasons, and eight QTLs for two traits were conserved across P levels. Low-P-specific QTLs were clustered on chromosomes A1, A6 and A8. By comparative mapping between Arabidopsis and B. napus, 161 orthologues of 146 genes involved in Arabidopsis P homeostasis and/or yield-related trait control were associated with 45 QTLs corresponding to 23 chromosomal regions. Four gene-based markers developed from genes involved in Arabidopsis P homeostasis were mapped to QTL intervals.

Conclusions

Different genetic determinants were involved in controlling seed yield and yield-related traits in B. napus under normal and low P conditions. The QTLs detected under reduced P supply may provide useful information for improving the seed yield of B. napus in soils with low P availability in marker-assisted selection.     

Dissecting Quantitative Trait Loci for Boron Efficiency across Multiple Environments in Brassica napus

High yield is the most important goal in crop breeding, and boron (B) is an essential micronutrient for plants. However, B deficiency, leading to yield decreases, is an agricultural problem worldwide. Brassica napus is one of the most sensitive crops to B deficiency, and considerable genotypic variation exists among different cultivars in response to B deficiency. To dissect the genetic basis of tolerance to B deficiency in B. napus, we carried out QTL analysis for seed yield and yield-related traits under low and normal B conditions using the double haploid population (TNDH) by two-year and the BQDH population by three-year field trials. In total, 80 putative QTLs and 42 epistatic interactions for seed yield, plant height, branch number, pod number, seed number, seed weight and B efficiency coefficient (BEC) were identified under low and normal B conditions, singly explaining 4.15–23.16% and 0.53–14.38% of the phenotypic variation. An additive effect of putative QTLs was a more important controlling factor than the additive-additive effect of epistatic interactions. Four QTL-by-environment interactions and 7 interactions between epistatic interactions and the environment contributed to 1.27–4.95% and 1.17–3.68% of the phenotypic variation, respectively. The chromosome region on A2 of SYLB-A2 for seed yield under low B condition and BEC-A2 for BEC in the two populations was equivalent to the region of a reported major QTL, BE1. The B. napus homologous genes of Bra020592 and Bra020595 mapped to the A2 region and were speculated to be candidate genes for B efficiency. These findings reveal the complex genetic basis of B efficiency in B. napus. They provide a basis for the fine mapping and cloning of the B efficiency genes and for breeding B-efficient cultivars by marker-assisted selection (MAS).     

Quantitative trait loci affecting seed mineral concentrations in Brassica napus grown with contrasting phosphorus supplies

MethodsA population of 124 recombinant inbred lines derived from a cross between P-inefficient ‘B104-2’ and P-efficient ‘Eyou Changjia’ was used for phenotypic investigation and QTL analysis. Two-year field trials were conducted with two P treatments. Concentrations of mineral elements (P, Ca, Mg, Fe, Zn, Cu and Mn) in seeds were determined and QTLs were identified by composite interval mapping.ConclusionsThe accumulation of mineral elements in seeds is controlled by multiple genes. Common physiological and molecular mechanisms could be involved in the accumulation of several mineral elements, and genes involved in these processes in B. napus are suggested. These results offer insights to the genetic basis of seed mineral accumulation across different P levels in B. napus.     

Identification of QTLs associated with potassium use efficiency and underlying candidate genes by whole-genome resequencing of two parental lines in Brassica napus

Breeding crops that acquire and/or utilize potassium (K) more effectively could reduce the use of K fertilizers. Sixteen traits affecting K use efficiency (KUE) at the seedling stage were investigated in a B. napus double haploid population grown at an optimal K supply (OK) and a low K supply (LK) in a hydroponic culture system. In total, 50 and 62 QTLs associated with these traits were identified at OK and LK, respectively. A total of 25 orthologues of 23 Arabidopsis genes regulating K transport were identified in the confidence intervals of nine QTLs impacting shoot dry weight at LK, and 22 of these showed variations in coding sequences and/or exhibited significant differences in mRNA abundances in roots at LK between the two parental lines. This study provided insights to the genetic basis of KUE in B. napus, which will accelerate the breeding of K-efficient rapeseed cultivars by marker-assisted selection.     

A High-Density Genetic Map Identifies a Novel Major QTL for Boron Efficiency in Oilseed Rape (Brassica napus L.)

Low boron (B) seriously limits the growth of oilseed rape (Brassica napus L.), a high B demand species that is sensitive to low B conditions. Significant genotypic variations in response to B deficiency have been observed among B. napus cultivars. To reveal the genetic basis for B efficiency in B. napus, quantitative trait loci (QTLs) for the plant growth traits, B uptake traits and the B efficiency coefficient (BEC) were analyzed using a doubled haploid (DH) population derived from a cross between a B-efficient parent, Qingyou 10, and a B-inefficient parent, Westar 10. A high-density genetic map was constructed based on single nucleotide polymorphisms (SNPs) assayed using Brassica 60 K Infinium BeadChip Array, simple sequence repeats (SSRs) and amplified fragment length polymorphisms (AFLPs). The linkage map covered a total length of 2139.5 cM, with 19 linkage groups (LGs) and an average distance of 1.6 cM between adjacent markers. Based on hydroponic evaluation of six B efficiency traits measured in three separate repeated trials, a total of 52 QTLs were identified, accounting for 6.14–46.27% of the phenotypic variation. A major QTL for BEC, qBEC-A3a, was co-located on A3 with other QTLs for plant growth and B uptake traits under low B stress. Using a subset of substitution lines, qBEC-A3a was validated and narrowed down to the interval between CNU384 and BnGMS436. The results of this study provide a novel major locus located on A3 for B efficiency in B. napus that will be suitable for fine mapping and marker-assisted selection breeding for B efficiency in B. napus.     

Detection and validation of novel QTL for shoot and root traits in barley (Hordeum vulgare L.)

Shoot and root attributes are essential for plant performance in agriculture. Here, we report detection and validation of quantitative trait loci (QTL) for shoot and root traits in 301 BC₂DH lines achieved by crossing cultivar Scarlett and wild barley accession ISR42-8. Phenotypic evaluations were made for six traits across 3 years under control and drought conditions. QTL analysis was performed using 371 DNA markers genotyped by different protocols, such as sequence repeats, diversity array technology as well as gene-specific markers. Marker by trait analysis revealed 33 QTL of which 15 and 18 QTL showed trait-improving effects of the exotic and elite alleles, respectively. Two major QTL for plant height (PH) were found on chromosome 2H (QPh.S42.2H) and 3H (QPh.S42.3H.b). The strongest QTL QSdw.S42.5H for increasing shoot dry weight was associated with an exotic allele on chromosome 5H. QTL QTkw.S42.1H underlie a novel exotic allele that improved thousand kernel weight. Seven QTL were associated with root dry weight of which at four loci introgression of exotic alleles enhanced traits values. The strongest QTL QRdw.S42.7H was linked to a gene-specific marker VrnH3 on chromosome 7H. At QRl.S42.5H, the exotic allele accounted for a 9 % increase in root length. In addition, 18 epistatic interactions were linked to PH, shoot and root dry weights. QTL validation was performed with 53 introgression lines (ILs) carrying ISR42-8 introgressions in the Scarlett background. Nine novel QTL alleles of exotic origin were validated in the isogenic background. These QTL-bearing ILs provide valuable genetic resources for plant breeding and positional cloning of the underlying genes.     

Phosphate-assisted phytoremediation of arsenic by Brassica napus and Brassica juncea: Morphological and physiological response

In this study, we examined the potential role of phosphate (P; 0, 50, 100 mg kg^?1) on growth, gas exchange attributes, and photosynthetic pigments of Brassica napus and Brassica juncea under arsenic (As) stress (0, 25, 50, 75 mg kg^?1) in a pot experiment. Results revealed that phosphate supplementation (P100) to As-stressed plants significantly increased shoot As concentration, dry biomass yield, and As uptake, in addition to the improved morphological and gas exchange attributes and photosynthetic pigments over P0. However, phosphate-assisted increase in As uptake was substantially (up to two times) greater for B. napus, notably due to higher shoot As concentration and dry biomass yield, compared to B. juncea at the P100 level. While phosphate addition in soil (P100) led to enhanced shoot As concentration in B. juncea, it reduced shoot dry biomass, primarily after 50 and 75 mg kg^?1 As treatments. The translocation factor and bioconcentration factor values of B. napus were higher than B. juncea for all As levels in the presence of phosphate. This study demonstrates that phosphate supplementation has a potential to improve As phytoextraction efficiency, predominantly for B. napus, by minimizing As-induced damage to plant growth, as well as by improving the physiological and photosynthetic attributes.     

小麦幼苗耐热性的QTL定位分析

以小麦DH群体(‘旱选10号’ב鲁麦14’)为材料,在高温(热胁迫)及常温(对照)两种条件下考察小麦幼苗的根干重、苗干重、幼苗生物量、叶片叶绿素含量、叶绿素荧光参数及其耐热指数,并应用基于混合线性模型的复合区间作图法分析幼苗性状及其耐热指数QTL的数量、染色体分布及表达情况,以及QTL与环境的互作效应。结果显示:(1)亲本‘旱选10号’的耐热性明显优于‘鲁麦14’,且杂交后代的耐热性出现超亲分离。(2)控制幼苗耐热相关性状的QTL位点在染色体2D、6B、3A、4A、5A和7A上分布较多,而控制幼苗性状耐热指数的QTL在染色体6A、6B、3A、2D、5A和7A上分布较多,QTL位点在染色体上的分布有区域化的趋势。(3)控制幼苗性状的单个加性QTL和上位性QTL解释的表型变异分别平均为2.48%和2.65%;而控制耐热指数的单个加性QTL和上位性QTL解释的表型变异分别平均为8.84%和1.98%。(4)在热胁迫和对照条件下共检测到与幼苗性状及其耐热指数有关的加性效应QTL 13个和上位性效应QTL 28对,分布在除4D和6D以外的19条染色体上。研究表明,控制幼苗性状的QTL以上位性效应为主,而其耐热指数的QTL以加性效应为主。     

Analysis of genetic factors that control shoot mineral concentrations in rapeseed (Brassica napus) in different boron environments

Mineral nutrients are essential for plant cell function, and understanding the genetic and physiological basis of mineral concentration is therefore important for the development of nutrient-efficient crop varieties that can cope with a shortage of mineral resources. In the present study, we investigated the profiles of B, Ca, Fe, Cu, Mg, P and Zn concentrations in shoots and analyzed the genetic variation in a rapeseed (Brassica napus) double haploid population at normal and deficient boron (B) levels in hydroponic conditions. Significant correlations between the concentrations of different minerals, such as Ca and Mg, Ca and P, and Cu and Fe, existed in both B environments. A total of 35 quantitative trait loci (QTL) and 74 epistatic interaction pairs for mineral concentrations were identified by whole genome analysis of QTL and epistatic interactions. The individual phenotypic contributions of the QTL ranged from 4.4% to 19.0%, and the total percentage of genetic variance that was due to QTL and epistatic interactions varied from 10.4% to 82.4%. Most of these QTL corresponded specifically to one of the two B conditions except for one stable main-effect P-QTL across the B environments. Three QTL for Ca and Mg were found to co-localize under normal B condition. These results revealed that genetic factors control mineral homeostasis in plants and multigenes involving ion transport are required to regulate mineral balance in plants under conditions of diverse nutrient stress. In addition, 26 genes involved in ion uptake and transport in Arabidopsis thaliana were in silico mapped onto the QTL intervals of B. napus by comparative genomic analysis. These candidate orthologous genes in B. napus allowed the selection of genes involved in the controlling mineral concentration that may account for the identified QTL.     

Multi-trait and multi-environment QTL analysis reveals the impact of seed colour on seed composition traits in <Emphasis Type="Italic">Brassica napus</Emphasis>

Brassica napus seed composition traits (fibre, protein, oil and fatty acid profiles), seed colour and yield-associated traits are regulated by a complex network of genetic factors. Although previous studies have attempted to dissect the underlying genetic basis for these traits, a more complete picture of the available quantitative trait loci (QTL) variation and any interaction between the different traits is required. In this study, QTL mapping for eleven seed composition traits, seed colour and a yield-related trait (TSW) was conducted in a spring-type canola-quality B. napus doubled haploid (DH) population from a cross between black-seeded (DH12075) and yellow-seeded (YN01-429) lines across five environments. A major QTL associated with fibre traits (acid detergent fibre, acid detergent lignin and neutral detergent fibre) and seed colour (whiteness index) was mapped on chromosome N9 across the five environments. Multi-trait analysis identified QTL which had pleiotropic effect for seed colour and other composition traits. Multi-environment analysis revealed genetic (QTL) × environment effects on most QTL. These findings provide a more detailed insight into the complex QTL networks controlling seed composition and yield-associated traits in canola-quality B. napus.     

QTL analyses of drought tolerance and growth for a Salix dasyclados × Salix viminalis hybrid in contrasting water regimes

Quantitative trait loci (QTL) for growth traits and water-use efficiency have been identified in two water regimes (normal and drought-treated) and for a treatment index. A tetraploid hybrid F₂ population originating from a cross between a Salix dasyclados clone (SW901290) and a Salix viminalis clone (Jorunn) was used in the study. The growth response of each individual including both above and below ground dry-matter production (i.e. shoot length, shoot diameter, aboveground and root dry weight, internode length, root dry weight/total dry weight, relative growth rate and leaf nitrogen content) was analysed in a replicated block experiment with two water treatments. A composite interval mapping approach was used to estimate number of QTL, the magnitude of the QTL and their position on genetic linkage maps. QTL specific for each treatment and for the treatment index were found, but QTL common across the treatments and the treatment index were also detected. Each QTL explained from 8% to 29% of the phenotypic variation, depending on trait and treatment. Clusters of QTL for different traits were mapped close to each other at several linkage groups, indicating either a common genetic base or tightly linked QTL. Common QTL identified between treatments and treatment index in the complex trait dry weight can be useful tools in the breeding and selection for drought stress tolerance in Salix.     

Root and shoot traits responses to phosphorus deficiency and QTL analysis at seedling stage using introgression lines of rice

Phosphorous （P） deficiency is a major restraint factor for crop production and plants have developed several mechanisms to adapt to low P stress. In this study, a set of 271 introgression lines （ILs） were used to characterize the responses of seedlings to low P availability and to identify QTLs for root traits, biomass, and plant height under P-deficiency and P-sufficiency conditions. Plant height, total dry weight, shoot dry weight, and root number were inhibited under P-deficiency, whereas maximum root length （MRL） and root-shoot ratio （RS） were induced by P-deficiency stress. Relative MRL （RMRL, the ratio of MRL under P-deficiency to MRL under P-sufficiency con- dition） and relative RS （RRS） were used to evaluate P-deficiency tolerance at the seedling stage. A total of 24 additive QTLs and 29 pairs of epistatic QTLs were detected, but only qRN4 was detected in both conditions. This suggested that different mechanisms may exist in both P supply levels. QTLs for adaptive traits （RMRL, RRS, RRV, and RRDW） and qRN4 consistently expressed to increase trait stability may contribute to P-deficiency tolerance. Twelve intervals were cluster regions of QTLs for P-deficiency tolerance, and one QTL （qRRSS） showed pleiotropic effects on P-deficiency tolerance and drought tolerance. These interesting QTLs can be used in marker-assisted breeding through the target ILs.     

Differential sensitivity of canola (Brassica napus) seedlings to ultraviolet-B radiation,water stress and abscisic acid

Responses of canola (Brassica napus L.) seedlings to three ultraviolet (UV)-B levels [0 (zero), 5 (ambient) and 10 (enhanced) kJ m^?2 d^?1], two watering regimes (well-watered and water-stressed), and two abscisic acid (ABA) levels (with and without application) were investigated. Overall, enhanced UVB and water stress negatively affected plant growth and physiology, but ABA had very little effect. Enhanced UVB decreased stem height, leaf area, plant dry matter, water use efficiency and wax content, but increased concentrations of chlorophyll a, carotenoids and flavonoids, and ethylene evolution. Water stress reduced stem height and diameter, leaf area, plant dry matter, leaf weight ratio and shoot:root weight ratio under zero and ambient UVB. Water stress also reduced chlorophyll a and carotenoids in plants exposed to enhanced UVB. ABA with watering regime had significant interactive effects only on leaf dry matter and wax content. We found that enhanced UVB and water stress adversely affected B. napus seedlings. Interaction between these two factors affected plant performance. In this interaction, ABA had little significant role. Also, optimum vegetative growth and biomass were achieved under ambient UVB.     

Identifying the limitations for growth in low performing piglets from birth until 10 weeks of age

The evolution of hyper-prolific pig breeds has led to a higher within-litter variation in birth weight and in BW gain during the nursery phase. Based on an algorithm developed in previous research, two populations from a pool of 368 clinically healthy piglets at 6 weeks of age were selected: a low (LP) and a high (HP) performing population and their development was monitored until the end of the nursery phase (10 weeks of age). To understand the cause of the variation in growth between these populations we characterized the LP and HP piglets in terms of body morphology, behaviour, voluntary feed intake, BW gain, and apparent total tract and ileal nutrient digestibility. Piglets were housed individually and were fed a highly digestible diet. At selection, 6 weeks of age, the BW of LP and HP piglets were 6.8±0.1 and 12.2±0.1 kg, respectively. Compared with the LP piglets the HP piglets grew faster (203 g/day), ate more (275 g/day) from 6 to 10 weeks of age and were heavier at 10 weeks (30.0 v. 18.8 kg, all P<0.01). Yet, the differences in average daily gain and average daily feed intake disappeared when compared per kg BW^0.75. Assuming similar maintenance requirements per kg BW^0.75 the efficiency of feed utilization above maintenance was 0.1 g/g lower for the LP piglets (P=0.09).The gain : feed ratio was similar for both groups. LP piglets tended to take more time to touch a novel object (P=0.10), and spent more time eating (P<0.05). At 10 weeks, LP piglets had a higher body length and head circumference relative to BW (P<0.01). Relative to BW, LP had a 21% higher small intestine weight; 36% longer length, and relative to average FI, the small intestinal weight was 4 g/kg higher (both P=<0.01). Apparent total tract and ileal dry matter, N and gross energy digestibility were similar between groups (P>0.10). We concluded that the low performance of the LP piglets was due to their inability to engage compensatory gain or compensatory feed intake as efficiency of nutrient utilization and feed intake per kg BW^0.75 was unaffected. LP piglets tend to be more fearful towards novel objects. The morphological comparisons, increased body length and head circumference relative to BW imply that LP piglets have an increased priority for skeletal growth.