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1.
A genetic linkage map of Salix (2n = 38), composed of 325 AFLP and 38 RFLP markers has been constructed. The map was based on a population ( n = 87) derived from a cross between the male hybrid clone "Bj?rn" ( Salix viminalis x Salix schwerinii) and the female clone "78183" ( S. viminalis). Three hundred fifty seven AFLPs corresponding to DNA polymorphisms heterozygous in one parent and null in the other were scored. A total of 87 RFLP probes, most (83) derived from the Populus genome, yielded 39 and 11 polymorphic loci segregating in a 1:1 and 1:2:1 ratio respectively. Two maps, one for each parent, were constructed according to the "two-way pseudo-testcross" mapping strategy. The S. viminalis x S. schwerinii map (2,404 cM) included 217 markers and formed 26 major linkage groups while S. viminalis (1,844 cM) consisted of 146 markers placed on 18 major groups. In addition, eight and 14 additional minor linkage groups composed of less than four markers (doubles and triplets) were obtained in the S. viminalis x S. schwerinii and the S. viminalis maps, respectively. Both maps provided 70-80% genome coverage with an average density of markers of 14 cM. To investigate possible homologies between the parental maps, 20 AFLPs and 11 RFLPs segregating in 3:1 or 1:2:1 ratios were included in the linkage analysis. Eight linkage groups homologous between the two maps were detected. The present genetic map was used to identify quantitative trait loci (QTLs) affecting growth-related traits. Eleven QTLs were identified; seven QTLs for height growth, one QTL for stem diameter, one QTL for the height: diameter ratio, one QTL for the number of vegetative buds during flowering time and one QTL for the number of shoots. The estimated magnitude of the QTL effect ranged from 14 to 22% of the total phenotypic variance. One QTL associated with height growth and one affecting the height: diameter ratio were overlapping in the same marker interval with the QTL affecting stem diameter. QTL stability over years was estimated for traits measured in multiple years. Generally, QTLs were only significant in a single year although two QTLs for height growth were close to reaching the significance level in 2 consecutive years.  相似文献   

2.
The genetic control of bud phenology in hybrid poplar was studied by mapping quantitative trait loci (QTL) affecting the timing of autumn bud set and spring bud flush. The founders of the mapping pedigree were collected from widely separated latitudes to maximize segregating variation for dormancy-related traits in the F(2) generation-the female Populus trichocarpa parent is from Washington State (48 degrees N) and the male P. deltoides parent is from Texas (31 degrees N). Bud set and bud flush timing were measured on the F(2) generation in a replicated clonal field trial. Using a linkage map constructed of AFLP and microsatellite markers, three QTL controlling bud set and six QTL controlling bud flush were detected. Additionally, five candidate genes believed to be involved in perception of photoperiod (PHYB1, PHYB2) or transduction of abscisic acid response signals (ABI1B, ABI1D, and ABI3) were placed on the QTL map. PHYB2 and ABI1B were found to be coincident with QTL affecting bud set and bud flush.  相似文献   

3.

Background

The genomic architecture of bud phenology and height growth remains poorly known in most forest trees. In non model species, QTL studies have shown limited application because most often QTL data could not be validated from one experiment to another. The aim of our study was to overcome this limitation by basing QTL detection on the construction of genetic maps highly-enriched in gene markers, and by assessing QTLs across pedigrees, years, and environments.

Results

Four saturated individual linkage maps representing two unrelated mapping populations of 260 and 500 clonally replicated progeny were assembled from 471 to 570 markers, including from 283 to 451 gene SNPs obtained using a multiplexed genotyping assay. Thence, a composite linkage map was assembled with 836 gene markers. For individual linkage maps, a total of 33 distinct quantitative trait loci (QTLs) were observed for bud flush, 52 for bud set, and 52 for height growth. For the composite map, the corresponding numbers of QTL clusters were 11, 13, and 10. About 20% of QTLs were replicated between the two mapping populations and nearly 50% revealed spatial and/or temporal stability. Three to four occurrences of overlapping QTLs between characters were noted, indicating regions with potential pleiotropic effects. Moreover, some of the genes involved in the QTLs were also underlined by recent genome scans or expression profile studies. Overall, the proportion of phenotypic variance explained by each QTL ranged from 3.0 to 16.4% for bud flush, from 2.7 to 22.2% for bud set, and from 2.5 to 10.5% for height growth. Up to 70% of the total character variance could be accounted for by QTLs for bud flush or bud set, and up to 59% for height growth.

Conclusions

This study provides a basic understanding of the genomic architecture related to bud flush, bud set, and height growth in a conifer species, and a useful indicator to compare with Angiosperms. It will serve as a basic reference to functional and association genetic studies of adaptation and growth in Picea taxa. The putative QTNs identified will be tested for associations in natural populations, with potential applications in molecular breeding and gene conservation programs. QTLs mapping consistently across years and environments could also be the most important targets for breeding, because they represent genomic regions that may be least affected by G × E interactions.  相似文献   

4.
Thirty three unique quantitative trait loci (QTLs) affecting the timing of spring bud flush have been identified in an intraspecific mapping population of coastal Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco var. menziesii]. Both terminal and lateral bud flush were measured over a 4-year period on clonal replicates at two test sites, allowing for the repeated estimation of QTLs. QTLs were detected on 12 linkage groups and, in general, each explained a small proportion of the total phenotypic variance and were additive in effect. Several QTLs influence the timing of bud flush over multiple years, supporting earlier evidence that the timing of bud flush through developmental stages is under moderate to strong genetic control by the same suite of genes through developmental stages. However, only a few QTLs controlling the timing of bud flush were detected at both test sites, suggesting that geographic location plays a major role in the phenology of spring growth. A small number of QTLs with year and site interactions were also estimated. Received: 20 July 2000 / Accepted: 19 October 2000  相似文献   

5.
Genetic variation of bud burst and early growth components was estimated in a full-sib family of Quercus robur L. comprising 278 offspring. The full sibs were vegetatively propagated, and phenotypic assessments were made in three field tests. This two-generation pedigree was also used to construct a genetic linkage map (12 linkage groups, 128 markers) and locate quantitative trait loci (QTLs) controlling bud burst and growth components. In each field test, the date of bud burst extended over a period of 20 days from the earliest to the latest clone. Bud burst exhibited higher heritability (0.15–0.51) than growth components (0.04–0.23) and also higher correlations across field tests. Over the three tests there were 32 independent detected QTLs (P5% at the chromosome level) controlling bud burst, which likely represent at least 12 unique genes or chromosomal regions controlling this trait. QTLs explained from 3% to 11% of the variance of the clonal means. The number of QTLs controlling height growth components was lower and varied between two and four. However the contribution of each QTL to the variance of the clonal mean was higher (from 4% to 19%). These results indicate that the genetic architecture of two important fitness-related traits are quite different. On the one hand, bud burst is controlled by several QTLs with rather low to moderate effects, but contributing to a high genetic (additive) variance. On the other hand, height growth depends on fewer QTLs with moderate to strong effects, resulting in lower heritabilities of the trait.  相似文献   

6.
Quantitative trait locus (QTL) analysis was performed at different time points during cold-acclimation of a tetraploid F2 Salix pedigree. The pedigree (n=92) was derived from a cross between a frost-susceptible diploid female clone Jorunn (Salix viminalis) and a frost resistant hexaploid male clone SW901290 (Salix dasyclados). Freezing resistance, height growth increment and number of new leaves were assessed at days 0, 12, 20, 24, 31 and 42 of a short day–low temperature (SD-LT) hardening regime, while the initiation of shoot tip abscission and shoot tip abscission were measured daily. Height increment, dry-to-fresh weight ratio and number of new leaves were also measured in a replicated field trial. Freezing resistance was determined from electrolyte leakage of leaf tissues and from visual injuries on stem segments, after exposure to a predetermined freeze-thaw stress. Using a genetic map of the F2 composed of 432 single-dose AFLP markers, a total of 19 genomic regions controlling freezing resistance (10) and phenological traits (9) before and during cold-acclimation (SD-LT) were identified. The magnitude of the phenotypic variation explained by each freezing resistance locus varied over acclimation time (0–45%), and there was no time point at which all the QTLs could be detected. The single QTL detected for non-acclimated freezing resistance did not reach significance at any time point during cold-acclimation, suggesting an independent genetic relationship between non-acclimated and acclimated resistance to freezing in Salix. Five of the loci associated with freezing resistance shared common intervals with loci controlling phenological traits. Of the 14 QTLs controlling autumn freezing resistance and/or phenological traits in the indoors experiment, six (43%) were associated with autumn phenology-related traits, i.e. height increment, dry-to-fresh weight ratio and number of new leaves, measured in the field. A major locus with multi-trait association in both indoor and outdoor experiments was detected.Communicated by O. Savolainen  相似文献   

7.
 Regions of the genome influencing height and leaf area in seedlings of a three-generation outbred pedigree of Eucalyptus nitens have been identified. Three QTLs affecting height and two QTLs affecting leaf area were located using single-factor analysis of variance. The three QTLs affecting height each explained between 10.3 and 14.7% of the phenotypic variance, while the two QTLs for leaf area each explained between 9.8 and 11.6% of the phenotypic variation. Analysis of fully informative marker loci linked to the QTLs enabled the mode of action of the QTLs to be investigated. For three loci the QTL effect segregated from only one parent, while for two loci the QTL showed multiple alleles and the effect segregated from both parents in the pedigree. The two QTLs affecting leaf area were located in the same regions as two of the QTLs affecting height. Analysis of these regions with fully informative markers showed that both QTLs were linked to the same markers, but one had a similar size of effects and a similar mode of action for both height and leaf area, whilst the other showed a different mode of action for the two traits. These regions may contain two closely linked genes or may involve a single gene with a pleiotrophic effect on both height and leaf area. The QTL with the greatest effect showed multiple alleles and an intra-locus interaction that reduced the size of the effect. Assessment for two of the QTLs in a second related family did not show an effect associated with the marker loci; however, this was consistent with the mode of action of these QTLs and the pattern of inheritance in the second family. Received: 1 August 1996 / Accepted: 25 October 1996  相似文献   

8.
Quantitative trait loci (QTLs) affecting fall and spring cold-hardiness were identified in a three-generation outbred pedigree of coastal Douglas-fir [Pseudotsuga meniziesii (Mirb.) Franco var. menziesii]. Eleven QTLs controlling fall cold-hardiness were detected on four linkage groups, and 15 QTLs controlling spring cold-hardiness were detected on four linkage groups. Only one linkage group contained QTLs for both spring and fall cold-hardiness, and these QTLs tended to map in close proximity to one another. Several QTLs were associated with hardiness in all three shoot tissues assayed in the spring, supporting previous reports that there is synchronization of plant tissues during de-acclimatization. For fall cold-hardiness, co-location of QTLs was not observed for the different tissues assayed, which is consistent with previous reports of less synchronization of hardening in the fall. In several cases, QTLs for spring or fall cold-hardiness mapped to the same location as QTLs controlling spring bud flush. QTL estimations, relative magnitudes of heritabilities, and genetic correlations based on clonal data in this single full-sib family, supports conclusions about the genetic control and relationships among cold-hardiness traits observed in population samples of Douglas-fir in previous studies. Received: 20 July 2000 / Accepted: 19 October 2000  相似文献   

9.
 Quantitative trait loci (QTLs) for annual height- and diameter-increment growth were estimated in each of two three-generation loblolly pine pedigrees. Annual height-increment was measured in three consecutive years in the BASE pedigree and in four consecutive years in the QTL pedigree. Annual diameter-increment was measured only in the QTL pedigree for two consecutive years. An interval mapping-approach was used to estimate the number of QTLs, the magnitude of QTL effects, and their position on genetic linkage maps. Thirteen different height-increment and eight different diameter-increment QTLs were detected, suggesting that the these traits are, at least in part, controlled by a few genes of large effect. Little evidence was found for the expression of individual QTLs in multiple years or in multiple genetic backgrounds. These results were discussed in terms of the power of the experiment and their consequences for marker-assisted breeding. Received: 12 July 1998/Accepted: 5 August 1998  相似文献   

10.
Random amplified polymorphic DNAs (RAPDs) were used in combination with a double pseudo-testcross mapping strategy to estimate the position and effects of quantitative trait loci (QTLs) for traits influencing juvenile tree growth and development in two apple cultivars. The mapping population consisted of 172 F1 trees from a cross between the columnar mutant ‘Wijcik McIntosh’ and a standard form disease-resistant selection NY 75441-58. Significant associations were found between markers and height increment, internode number, internode length, base diameter increment, base diameter after 9 years of growth, branch number, and leaf break. The number of genomic regions associated with each trait varied from one to eight. The amount of variation explained by linear regression on individual marker loci (R2) ranged from 3.9 to 24.3%, with an average of 7%. Multiple regression using markers for each putative QTL explained from 6.6 to 41.6% of the phenotypic variation, with an average value of 24.3%. A large number of traits had significant variation associated with the map position of the dominant columnar gene, Co. QTL stability over years was estimated by comparing the locations of putative QTLs for traits measured in multiple years. The majority of genomic regions were associated with a trait in only a single year, although regions associated with a trait in more than 1 year were also detected. The limitations of dominant markers and an outbred mapping pedigree for QTL analysis are discussed. Received: 27 August 1997 / Accepted: 10 February 1998  相似文献   

11.
Phenotypic correlations and quantitative trait loci (QTL) for important growth traits and a surrogate of intrinsic water-use efficiency (leaf delta(13)C) were analysed in a willow pedigree of 92 full-sibling clones grown under two water regimes. The major objective was to examine the genetic basis of the phenotypic correlations. Cuttings of Salix were glasshouse-grown during one growing season. The relative growth rate (RGR) and underlying traits were assessed. QTL analysis was conducted based on an available linkage map for Salix. Leaf area productivity and leaf nitrogen productivity were more important in determining RGR than leaf area ratio and specific leaf area. However, phenotypic correlations among growth traits partly varied between the two environments. QTL were detected for most growth traits, among them many common QTL for different traits. The QTL pattern reflected the phenotypic correlation pattern. None of the QTL for the complex traits was consistent across the different environments. The results demonstrate a genetic basis for phenotypic correlations among growth traits in Salix, and provide evidence for the existence of 'master switches' regulating some of the traits.  相似文献   

12.
云南元江普通野生稻株高和抽穗期QTL定位研究   总被引:5,自引:0,他引:5  
以云南元江普通野生稻为供体亲本,在特青的遗传背景下构建了一套BC3高代回交群体。利用117个SSR标记分析383个BC3F2株系的基因型,采用单标记分析法对控制元江普野株高和抽穗期的QTL进行分析。在北京和合肥两个地点试验结果表明,控制株高的QTL分布在第1染色体上,在RM104附近有一个QTL,与sd-1位置相当,其对表现型变异的贡献率在两个地点分别为27%和28%,其加性效应值分别为26.24cm和26.28cm,来自野生稻的等位基因显著提高回交群体的株高;在第1、3、7、8、11染色体共检测到6个控制抽穗期QTL,其中第8染色体RM25附近控制抽穗期的QTL在两个地点的贡献率分别为13%和15%,加性效应值为4.60d和3.65d,来自野生稻的等位基因使回交群体抽穗期延迟。  相似文献   

13.
 Quantitative traits, including juvenile growth, flower bearing and rooting ability, of a woody plant species, Cryptomeria japonica D. Don, were analyzed in a three-generation pedigree with 73 F2 progenies using a linkage map with 85 genetic markers (72 RFLP, 11 RAPD, one isozyme and one morphological loci). A cluster of quantitative trait loci (QTLs) related to juvenile growth and female flower bearing was detected on linkage group 2. Some of the influence of this cluster could be attributed to pleiotropic effects of a dwarf locus located in its vicinity. QTLs related to male and female flower bearing were detected at different locations and showed different effects from each other, suggesting that the genetic systems controlling male and female flowering are different. No large QTL affecting rooting ability was detected in the material analyzed in this study. Received: 15 December 1997 / Accepted: 4 February 1998  相似文献   

14.
Quantitative trait loci (QTLs) for body weight and tail length are mapped in an F2 population of 927 C57BL/6J × DBA/2J mice. We test the concordance between the locations of the mapped QTLs with those detected by changes of marker frequency under artificial selection in a previous experiment with the same base population. The directions of effects of the QTLs are generally in agreement, and in many cases significant QTLs are found in similar map positions, but there are also discrepancies between the two experiments. There are indications of age-specific QTL effects on growth. For body weight traits, the genetic variation in the F2 appears to result from many loci with relatively small effects. For tail length at 10 weeks, however, a single QTL on Chromosome (Chr) 1 with a peak LOD score of ∼33 contributes most of the genetic variation detected, changes the trait value by about 6%, and explains about 20% of the phenotypic variance of the trait. Received: 4 August 1998 / Accepted: 17 November 1998  相似文献   

15.
Fusarium head blight (FHB), an important disease of barley in many areas of the world, causes losses in grain yield and quality. Deoxynivalenol (DON) mycotoxin residues, produced by the primary pathogen Fusarium graminearum, pose potential health risks. Barley producers may not be able to profitably market FHB-infected barley, even though it has a low DON level. Three types of FHB resistance have been described in wheat: Type I (penetration), Type II (spread), and Type III (mycotoxin degradation). We describe putative measures of these three types of resistance in barley. In wheat, the three resistance mechanisms show quantitative inheritance. Accordingly, to study FHB resistance in barley, we used quantitative trait locus (QTL) mapping to determine the number, genome location, and effects of QTLs associated with Type-I and -II resistance and the concentration of DON in the grain. We also mapped QTLs for plant height, heading date, and morphological attributes of the inflorescence (seeds per inflorescence, inflorescence density, and lateral floret size). QTL analyses were based on a mapping population of F1-derived doubled-haploid (DH) lines from the cross of the two-rowed genotypes Gobernadora and CMB643, a linkage map constructed with RFLP marker loci, and field evaluations of the three types of FHB resistance performed in China, Mexico, and two environments in North Dakota, USA. Resistance QTLs were detected in six of the seven linkage groups. Alternate favorable alleles were found at the same loci when different inoculation techniques were used to measure Type-I resistance. The largest-effect resistance QTL (for Type-II resistance) was mapped in the centromeric region of chromosome 2. All but two of the resistance QTLs coincided with QTLs determining morphological attributes of the inflorescence and/or plant height. Additional experiments are needed to determine if these coincident QTLs are due to linkage or pleiotropy and to more clearly define the biological basis of the FHB resistance QTLs. Plant architecture should be considered in FHB resistance breeding efforts, particularly those directed at resistance QTL introgression and/or pyramiding. Received: 22 November 1998 / Accepted: 2 June 1999  相似文献   

16.
Interval mapping of quantitative trait loci (QTL) for 16 yield, agronomic and quality traits in potato was performed on a tetraploid full-sib family comprising 227 clones from a cross between processing clone 12601ab1 and table cultivar Stirling. Thirty-eight AFLP primer combinations and six SSRs provided 514 informative markers which formed a molecular marker map comprising 12 linkage groups (LGs) in 12601ab1 (nine with four homologous chromosomes) which were aligned with 12 in Stirling (11 with four homologous chromosomes), with four partial groups remaining in 12601ab1. Two LGs were identified unequivocally as chromosomes IV and V and eight others were tentatively assigned with chromosomes VII and X unidentified. All of the traits scored had moderately high heritabilities with 54–92% of the variation in clone means over 3 years and two replicates being due to genetic differences. A total of 39 QTLs were identified. A QTL for maturity was identified on chromosome V which explained 56% of the phenotypic variance, whereas the other QTLs individually explained between 5.4 and 16.5%. However, six QTLs were detected for after-cooking blackening and four for each of regularity of tuber shape, fry colour both after storage at 4 and 10°C and sprouting. Just two QTLs were found for each of yield, the two ‘overall’ scores, crop emergence, tuber size and common scab and just one QTL was detected for each of dry matter content, keeping quality, growth cracks and internal condition. The implications for practical potato breeding and for practical linkage and QTL analysis in autotetraploids are discussed.  相似文献   

17.
A multiple marker least squares approach is presented for the analysis of a single three-generation pedigree for quantitative trait locus (QTL) characterisation. It is an extension of the approach by Haley et?al. (1994) to the situation where grandparents cannot be assumed to be homozygous at QTLs for the trait of interest. The method is applied to the analysis of wood specific gravity in loblolly pine (Pinus taeda L.). Within a similar framework, a series of preliminary analyses are carried out, followed by a more detailed search of the genome for one or more QTLs. The preliminary analyses provide information about whether the contribution from each linkage group appears to be polygenic, localised to a small region (e.g. a single QTL) or oligogenic (i.e. several QTLs). Significance levels are obtained using a permutation test that uses the observed phenotypes and marker genotypes. The conclusion of these analyses is that in this pedigree single QTLs with very large effect on wood specific gravity do not appear to be segregating, although there is evidence for QTLs with small effect. Finally, in order to assess the potential power of this pedigree, we simulated QTLs within the framework of the actual marker data. As expected, QTL effects would need to be large to be reliably detected in this study, and the power to detect QTLs varies at different positions in the genome depending on the level of information in the local markers.  相似文献   

18.
 Phosphorus (P) deficiency of soils is a major yield-limiting factor in rice production. Increasing the P-deficiency tolerance of rice cultivars may represent a more cost-effective solution than relying on fertilizer application. The objective of this study was to identify putative QTLs for P-deficiency tolerance in rice, using 98 backcross inbred lines derived from a japonica×indica cross and genotyped at 245 RFLP marker loci. Lines were grown on P-deficient soil and P uptake, internal P-use efficiency, dry weight, and tiller number were determined. Three QTLs were identified for dry weight and four QTLs for P uptake, together explaining 45.4% and 54.5% of the variation for the respective traits. Peaks for both traits were in good agreement which was to be expected considering the tight correlation of r=0.96 between dry weight and P uptake. For both traits the QTL linked to marker C443 on chromosome 12 had a major effect. Two of the three QTLs detected for internal P-use efficiency, including the major one on chromosome 12, coincided with QTLs for P uptake; however, whereas indica alleles increased P uptake they reduced P-use efficiency. We concluded that this was not due to the tight linkage of two genes in repulsion but rather due to an indirect effect of P uptake on P-use efficiency. Most lines with high use efficiency were characterized by very low P uptake and dry weight and apparently experienced extreme P-deficiency stress. Their higher P-use efficiency was thus the result of highly sub-optimal tissue-P concentrations and did not represent a positive adaptation to low P availability. The number of tillers produced under P deficiency is viewed as an indirect indicator of P-deficiency tolerance in rice. In addition to the major QTL on chromosome 12 already identified for all other traits, two QTLs on chromosome 4 and 12 were identified for tiller number. Their position, however, coincided with QTLs for tiller number reported elsewhere under P-sufficient conditions and therefore appear to be not related to P-deficiency tolerance. In this study P-deficiency tolerance was mainly caused by differences in P uptake and not in P-use efficiency. Using a trait indirectly related to P-deficiency tolerance such as tiller number, we detected a major QTL but none of the minor QTLs detected for P uptake or dry weight. Received: 9 February 1998 / Accepted: 29 April 1998  相似文献   

19.
Growth trajectories are a biological process important to plant and animal breeding, and to evolutionary genetic studies. In this article, we report the detection of quantitative trait loci (QTLs) responsible for growth trajectories in poplars that are used as a model system for the study of forest biology. These QTLs were localized on a genetic linkage map of polymorphic markers using a statistical mapping method incorporating growth-curve models. The effects of the QTLs on growth are described as a function of age, so that age-specific changes in QTL effects can be readily projected throughout the entire growth process. The QTLs identified display increased effects on growth when trees age, yet the timing of QTL activation is earlier for stem height than diameter, which is consistent with the ecological viewpoint of canopy competition. The implications of the results for breeding and silviculture are discussed.  相似文献   

20.
Tea plant (Camellia sinensis) is a major beverage crop across the world. To uncover the genetic controls of agronomic traits and facilitate marker-assisted breeding (MAB) in tea plant, we constructed a saturated SSR-based linkage map using an F1 population derived from the crossing of ‘Longjin43’ × ‘Baihaozao’. A total of 483 SSR markers, consisting of 117 novel loci, 129 transferred from other tea plant maps, and 237 previously mapped, were successfully integrated into a new consensus map. The map has 15 linkage groups, covering 1226.2 cM in total with an average marker distance of 2.5 cM. The 126 markers in common enabled us to align this map to the reference genetic maps of tea plant. Phenotype data were collected in 2014 and 2015 for five traits: timing of spring bud flush (TBF), young shoot color (YSC), mature leaf length (MLL), mature leaf width (MLW), and leaf shape index (LSI, i.e., MLL/MLW). QTL analyses were performed for the five traits using the new consensus map and 15 QTLs were identified. The SSR markers, linkage map, and QTLs reported here are useful resources for future QTL mining, identification of causal genes, and MAB in tea plant.  相似文献   

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