Genetic mapping in sugarcane, a high polyploid, using bi-parental progeny: identification of a gene controlling stalk colour and a new rust resistance gene

Modern sugarcane cultivars (Saccharum spp) are highly polyploïd and aneuploid interspecific hybrids (2n=100–130). Two genetic maps were constructed using a population of 198 progeny from a cross between R570, a modern cultivar, and MQ76-53, an old Australian clone derived from a cross between Trojan (a modern cultivar) and SES528 (a wild Saccharum spontaneum clone). A total of 1,666 polymorphic markers were produced using 37 AFLP primer combinations, 46 SSRs and 9 RFLP probes. Linkage analysis led to the construction of 86 cosegregation groups for R570 and 105 cosegregation groups for MQ76-53 encompassing 424 and 536 single dose markers, respectively. The cumulative length of the R570 map was 3,144 cM, while that of the MQ76-53 map was 4,329 cM. Here, we integrated mapping information obtained on R570 in this study with that derived from a previous map based on a selfed R570 population. Two new genes controlling Mendelian traits were localized on the MQ76-53 map: a gene controlling the red stalk colour was linked at 6.5 cM to an AFLP marker and a new brown rust resistance gene was linked at 23 cM to an AFLP marker. Besides another previously identified brown rust resistance gene (Bru1), these two genes are the only other major genes to be identified in sugarcane so far.     

Molecular insights into the origin of the brown rust resistance gene <Emphasis Type="Italic">Bru</Emphasis>1 among <Emphasis Type="Italic">Saccharum</Emphasis> species

Key message

Analysis of 387 sugarcane clones using Bru 1 diagnostic markers revealed two possible sources of Bru 1 in Chinese cultivars: one from Saccharum spontaneum and another from Saccharum robustum of New Guinea.

Abstract

Sugarcane brown rust (SBR) is an important fungal disease in many sugarcane production areas around the world, and can cause considerable yield losses in susceptible sugarcane cultivars. One major SBR resistance gene, named Bru1, initially identified from cultivar R570, was shown to be a major SBR resistance source in most of the sugarcane producing areas of the world. In this study, by using the two Bru1-associated markers, R12H16 and 9O20-F4, we surveyed the presence of Bru1 in a Chinese sugarcane germplasm collection of 387 clones, consisting of 228 hybrid cultivars bred by different Chinese sugarcane breeding establishments, 54 exotic hybrid cultivars introduced from other countries and 105 clones of sugarcane ancestral species. The Bru1-bearing haplotype was detected in 43.4% of Chinese sugarcane cultivars, 20.4% of exotic hybrid cultivars, and only 3.8% of ancestral species. Among the 33 Chinese cultivars for which phenotypes of resistance to SBR were available, Bru1 was present in 69.2% (18/26) of the resistant clones. Analyses of the allelic sequence variations of R12H16 and 9O20-F4 suggested two possible sources of Bru1 in Chinese cultivars: one from S. spontaneum and another from S. robustum of New Guinea. In addition, we developed an improved Bru1 diagnostic marker, 9O20-F4-HaeIII, which can eliminate all the false results of 9O20-F4-RsaI observed among S. spontaneum, as well as a new dominant Bru1 diagnostic marker, R12E03-2, from the BAC ShCIR12E03. Our results provide valuable information for further efforts of breeding SBR-resistant varieties, searching new SBR resistance sources and cloning of Bru1 in sugarcane.

     

Targeted mapping of a sugarcane rust resistance gene (Bru1) using bulked segregant analysis and AFLP markers

The presence of a major resistance gene (Bru1) for brown rust in the sugarcane cultivar R570 (2n about 115) was confirmed by analyzing segregation of rust resistance in a large population of 658 individuals, derived from selfing of clone R570. A subset of this population was analyzed with AFLP and bulked segregant analysis (BSA) to develop a detailed genetic map around the resistance gene. Four hundred and forty three primer pairs were used resulting in the identification of eight AFLP markers surrounding the resistance gene in an interval of 10 cM, with the closest markers located at 1.9 and 2.2 cM on each side of the gene. Efficiency of the AFLP/BSA applied to the complex polyploid genome of sugarcane is discussed, as well as the potential of the newly identified AFLP markers for developing a map-based cloning approach exploiting, synteny conservation with sorghum.Communicated by H. F. Linskens     

Diploid/polyploid syntenic shuttle mapping and haplotype-specific chromosome walking toward a rust resistance gene (Bru1) in highly polyploid sugarcane (2n approximately 12x approximately 115)

The genome of modern sugarcane cultivars is highly polyploid ( approximately 12x), aneuploid, of interspecific origin, and contains 10 Gb of DNA. Its size and complexity represent a major challenge for the isolation of agronomically important genes. Here we report on the first attempt to isolate a gene from sugarcane by map-based cloning, targeting a durable major rust resistance gene (Bru1). We describe the genomic strategies that we have developed to overcome constraints associated with high polyploidy in the successive steps of map-based cloning approaches, including diploid/polyploid syntenic shuttle mapping with two model diploid species (sorghum and rice) and haplotype-specific chromosome walking. Their applications allowed us (i) to develop a high-resolution map including markers at 0.28 and 0.14 cM on both sides and 13 markers cosegregating with Bru1 and (ii) to develop a physical map of the target haplotype that still includes two gaps at this stage due to the discovery of an insertion specific to this haplotype. These approaches will pave the way for the development of future map-based cloning approaches for sugarcane and other complex polyploid species.     

Utilization of a major brown rust resistance gene in sugarcane breeding

Brown rust, caused by Puccinia melanocephala, has had devastating effects on sugarcane (Saccharum spp.) breeding programs and commercial production. The discovery of Bru1, a major gene conferring resistance to brown rust, represented a substantial breakthrough. Markers for Bru1 are the first available for sugarcane molecular breeding. The contribution of Bru1 towards brown rust resistance in the Canal Point (CP) sugarcane breeding program was determined as a means of directing future breeding strategies. Bru1 was detected in 285 of 1,072 (27 %) clones used for crossing; this germplasm represents the genetic base for cultivar development in Florida. The frequency of Bru1 was greatest in CP clones (42 %) and lowest among Louisiana clones (6 %). Bru1 was not detected in clones with year assignments before 1953. However, Bru1 frequency increased from 15 % (assignments 1975–1985) to 47 % in the current decade. The increase coincided with the introduction of brown rust to Florida. Bru1 was detected in 155 (32 %) of 485 parental clones tested for brown rust susceptibility at two field locations. Of clones classed resistant to brown rust, 154 (59 %) contained Bru1, yet none of 100 susceptible clones contained the gene. Bru1 was detected in 667 (44 %) clones in the second clonal stage of selection, 87 % of which were free of brown rust symptoms. Bru1 is the predominant source of resistance in the Florida sugarcane genetic base. Efforts to identify and integrate new brown rust resistance genes must be pursued to minimize risks associated with a future breakdown in major gene resistance provided by Bru1.     

Identification and Validation of Molecular Markers Associated with Pachymetra Root Rot and Brown Rust Resistance in Sugarcane Using Map- and Association-based Approaches

Marker-assisted selection for traits that are difficult to screen for, such as resistance to many sugarcane diseases, has the potential to facilitate the development of improved cultivars in sugarcane. Pachymetra root rot (PRR) and brown rust resistance ratings were obtained over two years for 192 I1 progeny (progeny produced by two heterozygous, non-inbred parental lines) of a sugarcane (Saccharum spp. hybrid) cross between two elite sugarcane clones, Q117 and 74C42. Approximately 1000 single-dose markers, including microsatellite (SSR), amplified fragment length polymorphism (AFLP) and restriction fragment length polymorphism (RFLP) markers, were scored across the population and maps containing approximately 400 markers were constructed for each parent. At p ≤ 0.01, two genomic regions, one from the female Q117 map and a different region from the 74C42 male map, plus an unlinked bi-parental simplex marker (single-dose marker present in both parents) were identified as associated with PRR over both years of data collection. These regions explained between 6 and 16% of the phenotypic variation. An additional region was identified in the female map as associated with PRR at p ≤ 0.01 in one year and p ≤ 0.05 in the second year. This region explained between 4 and 8% of the phenotypic variation. For brown rust, two genomic regions, one from the female map and one from the male map, plus an unlinked marker from both maps, were identified as associated with brown rust resistance at p ≤ 0.01 over two years of phenotypic data. Each region explained between 7 and 18% of the phenotypic variation. Several additional regions were identified in both maps as associated with brown rust at p ≤ 0.01 in one year and p ≤ 0.05 in the second year. These regions also explained between 5 and 11% of the phenotypic variation. To validate these markers and determine whether they would be useful in alternative germplasm, markers from each genomic region associated with PRR or brown rust were screened across a set of 154 elite sugarcane clones; PRR and brown rust ratings were available for 131 and 72 of the clones, respectively. For PRR, three of the 6 markers tested remained significantly associated (p ≤ 0.01) with resistance ratings in the elite clone set. For brown rust, only one of the seven markers tested remained significantly associated (p ≤ 0.01) with resistance in the elite clone set, with one other marker associated at p ≤ 0.05. These results suggest that these markers could be broadly effective in selecting for PRR and/or brown rust resistance in sugarcane breeding programs.     

Quantitative trait loci for sugarcane resistance to the spotted stem borer <Emphasis Type="Italic">Chilo sacchariphagus</Emphasis>

The spotted stem borer (SSB) Chilo sacchariphagus is a major pest of sugarcane, causing substantial losses in cane weight and in sucrose yield. SSB resistance is an important trait to be taken into account for sugarcane breeding programs. In order to analyse the genetic basis of the resistance to SSB, we undertook a quantitative trait allele (QTA) mapping study based on a population of 147 progenies derived from the selfing of the resistant modern cultivar R570. The experimental population was evaluated in a replicated trial for borer damage under natural infestation in two successive crop cycles. A single-factor analysis using 1,405 polymorphic markers was performed to detect marker–trait associations. Statistical thresholds based on permutation tests designed to control type I errors at a low level allowed the detection of nine QTAs whose individual size ranged between 6 and 10% of the total variation. These nine QTAs are distributed over five of the eight homeology groups of the polyploid R570 genome. Two QTAs were found to co-localize with two typical resistance gene analog clusters. Overall, eight QTAs explain altogether 42% of the total phenotypic variance.     

Application of synteny across Poaceae to determine the map location of a sugarcane rust resistance gene

A major rust resistance gene has been identified in a self-progeny of the sugarcane cultivar R570. Until now, this gene was known to be linked to a marker revealed by the sugarcane probe CDSR29 but unassigned to any linkage group of the current genetic map. We used synteny relationships between sugarcane and three other grasses in an attempt to saturate the region around this rust resistance gene. Comparison of sugarcane, sorghum, maize and rice genetic maps led to the identification of homoeologous chromosome segments at the extremity of sorghum linkage group D, rice linkage group 2, maize linkage group 4 and in the centromeric region of maize linkage group 5. One hundred and eighty-four heterologous probes were selected and tested for cross-hybridization with sugarcane DNA; 106 produced a good hybridization signal and were hybridized on 88 individuals of the R570 selfed progeny. Two hundred and seventeen single-dose markers were added to the R570 genetic map, of which 66% mapped to linkage group VII, together with the rust resistance gene. This gene has now been mapped to the end of a co-segregating group consisting of 19 RFLP markers. None of the mapped loci were located closer to the gene than CDSR29. The gene thus appears to reside at the edge of a ’’synteny cluster’’ used to describe the different grass genomes. Received: 12 January 2000 / Accepted: 21 March 2000     

Constructing high-density genetic maps for polyploid sugarcane (<Emphasis Type="Italic">Saccharum</Emphasis> spp.) and identifying quantitative trait loci controlling brown rust resistance

Sugarcane (Saccharum spp.) is an important economic crop for producing edible sugar and bioethanol. Brown rust has long been a major disease impacting sugarcane production worldwide. Resistance resource and markers linked to resistance are valuable tools for disease resistance improvement. An F₁ segregating population derived from a cross between two hybrid sugarcane clones, brown rust-susceptible CP95-1039 and brown rust-resistant CP88-1762, were genotyped using genotyping by sequencing approach and also phenotyped in a replicated field trial. Single nucleotide polymorphism (SNP) and presence/absence markers were called with seven different pipelines to maximize reliable marker identification. High-density maps were constructed for both parental clones with a total map length of 4224.4 cM, and a marker density of one marker per 1.7 cM for CP95-1039, and a total map length of 4373.2 cM, and one marker per 2.0 cM for CP88-1762. Among the seven SNP callers, Tassel and Genome Analysis ToolKit performed better than other callers in single-dose SNP detection and contribution to genetic maps. Two major quantitative trait loci (QTL) controlling brown rust resistance were identified, which can explain 21 and 30% of the phenotypic variation, respectively. The genetic maps generated here will improve our understanding of sugarcane’s complex genome structure and discovery of underlying sequence variations controlling agronomic traits. The putative QTL controlling brown rust resistance can effectively be utilized in sugarcane breeding programs to expedite the selection process of brown rust resistance after validation.     

Assessment of genetic diversity of wheat genotypes by resistance gene analog-EST markers

Resistance gene analog-expressed sequence tag (RGA-EST)-based markers have been used for variety discrimination and studies of genetic diversity in wheat. Our aim is to increase the competitiveness of public wheat breeding programs through intensive use of modern selection technologies, mainly marker-assisted selection. The genetic diversity of 77 wheat nucleotide binding site (NBS)-containing RGA-ESTs was assessed. Resistant and susceptible bread wheat (Triticum aestivum) genotypes were used as sources of DNA for PCR amplifications. In our previous studies, the F? individuals derived from the combinations PI178383 x Harmankaya99, Izgi2001 x ES14, and Sonmez2001 x Aytin98 were evaluated for yellow rust resistance at both seedling and adult stages to identify DNA markers. We have now examined the genetic variability among the resistant and susceptible Turkish wheat cultivars for yellow rust disease and the mean genetic distance between the cultivars. The highest similarity was 0.500 between Harmankaya99 and Sonmez2001. The lowest similarity was 0.286 between Aytin98, PI178383 and Aytin98, ES14. A relatively high level (49.5%) of polymorphism was observed with 77 RGA-EST primers across the six wheat genotypes, despite the fact that all of them were local cultivars from geographically close locations. RGA-EST sequences were compared by BlastX algorithms for amino acid sequences to determine the polymorphic categories among the combinations. BlastX analyses of six RGA-ESTs that gave polymorphic patterns for all combinations were NBS-LRR class RGA, NB-ARC domain containing protein, NBS-type resistance protein RGC5, NBS-LRR-S/ TPK stem rust resistance protein, and putative MLA1 proteins, while 38 RGA-EST gave a monomorphic pattern.     

基因枪法介导GNA基因遗传转化甘蔗的研究

目的:将含有雪花莲外源凝集素(GNA)基因的植物表达载体用基因枪法分别导入一个果蔗和一个糖蔗品种中,以期获得转基因植株。方法:将GNA基因插入到植物表达载体上,构建出不同选择标记、不同启动子的表达载体,并用基因枪法将之导入甘蔗胚性愈伤组织,分别在G418、PPT和Hyg的选择压力下,筛选抗性植株,并进行分子杂交鉴定。结果:通过斑点杂交和PCR-Southern杂交证明GNA基因已整合到甘蔗基因组中。结论:用基因枪法成功获得了含有GNA基因的甘蔗转化株,为培育抗甘蔗绵蚜(Ceratovacuna lanigeraZehnther)的新品种提供了基础。     

Assessing wheat (Triticum aestivum) genotypes for "Yr" resistance genes using conserved regions and simple-sequence motifs

Analysis of DNA sequence variation among genotypes is useful for differentiation of wheat accessions, selection strategies and genetic development of crop plants. We screened molecular markers for yellow rust resistance genes (Yr7, Yr9, Yr15, Yr18, Yr26, and YrH52), which are in the gene-rich regions of wheat chromosomes 1B, 2B, and 7D, to investigate DNA sequence differences and repeat motifs and numbers between wheat cultivars resistant (Izgi2001, Sonmez2001, PI178383) and susceptible (Aytin98, ES14, Harmankaya99) to yellow rust. The F(2) individuals derived from the crosses were evaluated for yellow rust resistance at both the seedling and adult stages to identify DNA markers genetically linked to yellow rust resistance. The most repeated motif was found to be GA and the least repeated motif TAGA among the cultivars. When we examined DNA sequence differences (insertion, deletion and single nucleotide changes), the molecular markers Xgwm526 (Yr7) and Xgwm273 (YrH52) were found to have the most conserved regions and Yr15 (Xgwm413) the least conserved regions among the cultivars. This DNA sequence information can be used for selection of suitable parents, creating mapping populations and developing molecular markers associated with yellow rust resistance in plant breeding programs.     

Gene-specific markers for the wheat gene Lr34/Yr18/Pm38 which confers resistance to multiple fungal pathogens

The locus Lr34/Yr18/Pm38 confers partial and durable resistance against the devastating fungal pathogens leaf rust, stripe rust, and powdery mildew. In previous studies, this broad-spectrum resistance was shown to be controlled by a single gene which encodes a putative ATP-binding cassette transporter. Alleles of resistant and susceptible cultivars differed by only three sequence polymorphisms and the same resistance haplotype was found in the three independent breeding lineages of Lr34/Yr18/Pm38. Hence, we used these conserved sequence polymorphisms as templates to develop diagnostic molecular markers that will assist selection for durable multi-pathogen resistance in breeding programs. Five allele-specific markers (cssfr1–cssfr5) were developed based on a 3 bp deletion in exon 11 of the Lr34-gene, and one marker (cssfr6) was derived from a single nucleotide polymorphism in exon 12. Validation of reference genotypes, well characterized for the presence or absence of the Lr34/Yr18/Pm38 resistance locus, demonstrated perfect diagnostic values for the newly developed markers. By testing the new markers on a larger set of wheat cultivars, a third Lr34 haplotype, not described so far, was discovered in some European winter wheat and spelt material. Some cultivars with uncertain Lr34 status were re-assessed using the newly derived markers. Unambiguous identification of the Lr34 gene aided by the new markers has revealed that some wheat cultivars incorrectly postulated as having Lr34 may possess as yet uncharacterised loci for adult plant leaf and stripe rust resistance. E. S. Lagudah and S. G. Krattinger contributed equally to the work.     

河南省16个主栽小麦品种抗叶锈基因分析

为了明确河南省小麦品种的抗叶锈性及抗叶锈基因的分布,为小麦品种推广与合理布局、叶锈病防治及抗病育种提供依据,本研究利用2015年采自河南省的5个小麦叶锈菌流行小种混合菌株,对近几年河南省16个主栽小麦品种进行了苗期抗性鉴定,然后选用12个小麦叶锈菌生理小种对这些品种进行苗期基因推导,同时利用与24个小麦抗叶锈基因紧密连锁(或共分离)的30个分子标记对该16个品种进行了抗叶锈基因分子检测。结果显示,供试品种苗期对小麦叶锈菌混合流行小种均表现高度感病;基因推导与分子检测结果表明,供试品种可能含有Lr1、Lr16、Lr26和Lr30这4个抗叶锈基因,其中先麦8号含有Lr1和Lr26;郑麦366和郑麦9023含有Lr1;西农979和怀川916含有Lr16;中麦895、偃展4110、郑麦7698、平安8号、众麦1号、周麦16、衡观35和矮抗58含有Lr26;周麦22中含有Lr26,还可能含有Lr1和Lr30;豫麦49-198和洛麦23可能含有本研究中检测以外的其他抗叶锈基因。因此,河南省主栽小麦品种的抗叶锈基因丰富度较低,今后育种工作应注重引入其他抗叶锈性基因,提高抗叶锈性,有效控制小麦叶锈病。     

Resistance gene analogues in sugarcane and sorghum and their association with quantitative trait loci for rust resistance.

Fifty-four different sugarcane resistance gene analogue (RGA) sequences were isolated, characterized, and used to identify molecular markers linked to major disease-resistance loci in sugarcane. Ten RGAs were identified from a sugarcane stem expressed sequence tag (EST) library; the remaining 44 were isolated from sugarcane stem, leaf, and root tissue using primers designed to conserved RGA motifs. The map location of 31 of the RGAs was determined in sugarcane and compared with the location of quantitative trait loci (QTL) for brown rust resistance. After 2 years of phenotyping, 3 RGAs were shown to generate markers that were significantly associated with resistance to this disease. To assist in the understanding of the complex genetic structure of sugarcane, 17 of the 31 RGAs were also mapped in sorghum. Comparative mapping between sugarcane and sorghum revealed syntenic localization of several RGA clusters. The 3 brown rust associated RGAs were shown to map to the same linkage group (LG) in sorghum with 2 mapping to one region and the third to a region previously shown to contain a major rust-resistance QTL in sorghum. These results illustrate the value of using RGAs for the identification of markers linked to disease resistance loci and the value of simultaneous mapping in sugarcane and sorghum.     

Genomic distribution and characterization of EST-derived resistance gene analogs (RGAs) in sugarcane

A large sugarcane EST (expressed sequence tag) project recently gave us access to 261,609 EST sequences from sugarcane, assembled into 81,223 clusters. Among these, we identified 88 resistance gene analogs (RGAs) based on their homology to typical pathogen resistance genes, using a stringent BLAST search with a threshold e-value of e(-50). They included representatives of the three major groups of resistance genes with NBS/LRR, LRR or S/T KINASE domains. Fifty RGAs showed a total of 148 single-dose polymorphic RFLP markers, which could be located on the sugarcane reference genetic map (constructed in cultivar R570, 2n=approximately 115). Fifty-five SSR loci corresponding to 134 markers in R570 were also mapped to enable the classification of the various haplotypes into homology groups. Several RGA clusters were found. One cluster of two LRR-like loci mapped close to the only disease resistance gene known so far in sugarcane, which confers resistance to common rust. Detailed sequence comparison between two NBS/LRR RGA clusters in relation to their orthologs in rice and maize suggests their polyphyletic origins, and indicates that the degree of divergence between paralogous RGAs in sugarcane can be larger than that from an ortholog in a distant species.     

黄淮麦区小麦品种(系)中Yr26基因的SSR检测

选用与Yr26紧密连锁的SSR标记Xgwm11和Xgwm18结合田间抗性鉴定,对239份黄淮麦区小麦品种(系)进行检测,以明确Yr26基因在黄淮麦区小麦品种资源中的分布.结果表明:共有35份品种(系)含有与Yr26紧密连锁的SSR标记Xgwm18或Xgwm11的特征带,占检测样本的14.6%.在这35份材料中,31份田间抗性鉴定表现免疫至中抗,4份表现中感.分子标记检测与田间抗病性检测吻合度较好,该标记可以用于Yr26基因的分子标记辅助选择.综合分子标记和田间鉴定,31份小麦(系)含有Yr26基因,占102份抗病材料的30.39%.     

山东省12个主栽小麦品种(系)抗叶锈性分析

本研究旨在明确山东省12个小麦主栽品种(系)抗叶锈性及抗叶锈基因,为小麦品种推广与合理布局、叶锈病防治及抗病育种提供依据。利用2015年采自山东省的5个小麦叶锈菌流行小种的混合小种对这些材料进行苗期抗性鉴定,然后选用15个小麦叶锈菌生理小种对这些品种(系)进行苗期基因推导,并利用与24个小麦抗叶锈基因紧密连锁(或共分离)的30个分子标记对其进行抗叶锈基因分子检测。结果显示,山东省12个主栽小麦品种(系)苗期对该省2015年的5个小麦叶锈菌混合流行小种均表现高度感病。通过基因推导与分子检测发现,济南17含有Lr16,矮抗58和山农20含有Lr26,其余济麦系列、烟农系列、良星系列等9个品种(系)均未检测到所供试标记片段。此外,本研究还对山东省3个非主栽品种进行了检测,结果发现,中麦175含有抗叶锈基因Lr1和Lr37,含有成株抗性基因;皖麦38只检测到Lr26,济麦20未检测到所供试标记片段。综合以上结果,山东省主栽小麦品种(系)所含抗叶锈基因丰富度较低,尤其不含有对我国小麦叶锈菌流行小种有效的抗锈基因,应该引起高度重视,今后育种工作应注重引入其他抗叶锈基因,提高抗叶锈性。     

Exploiting a wheat EST database to assess genetic diversity

Expressed sequence tag (EST) markers have been used to assess variety and genetic diversity in wheat (Triticum aestivum). In this study, 1549 ESTs from wheat infested with yellow rust were used to examine the genetic diversity of six susceptible and resistant wheat cultivars. The aim of using these cultivars was to improve the competitiveness of public wheat breeding programs through the intensive use of modern, particularly marker-assisted, selection technologies. The F₂ individuals derived from cultivar crosses were screened for resistance to yellow rust at the seedling stage in greenhouses and adult stage in the field to identify DNA markers genetically linked to resistance. Five hundred and sixty ESTs were assembled into 136 contigs and 989 singletons. BlastX search results showed that 39 (29%) contigs and 96 (10%) singletons were homologous to wheat genes. The database-matched contigs and singletons were assigned to eight functional groups related to protein synthesis, photosynthesis, metabolism and energy, stress proteins, transporter proteins, protein breakdown and recycling, cell growth and division and reactive oxygen scavengers. PCR analyses with primers based on the contigs and singletons showed that the most polymorphic functional categories were photosynthesis (contigs) and metabolism and energy (singletons). EST analysis revealed considerable genetic variability among the Turkish wheat cultivars resistant and susceptible to yellow rust disease and allowed calculation of the mean genetic distance between cultivars, with the greatest similarity (0.725) being between Harmankaya99 and Sönmez2001, and the lowest (0.622) between Aytin98 and Izgi01.     

割手密野生资源抗逆性研究进展

割手密作为现代甘蔗遗传杂交育种史上最为成功的野生亲本,对多种不良环境都具有很强的抗逆性,被公认为是抗逆基因的主要来源。但目前真正被有效利用的割手密抗逆亲本和抗逆基因非常有限,我国自育和引进甘蔗主栽品种的抗逆性仍然比较单一且普遍偏弱,因此加强割手密优良抗逆亲本筛选和抗逆基因挖掘利用研究意义重大。本文综述了不同基因型割手密在非生物逆境(干旱、低温等理化因素)和生物逆境(病虫害侵染)下的抗逆性鉴定及其抗逆基因克隆和功能验证等国内外相关研究进展;并探讨了当前割手密资源抗逆材料筛选和抗逆基因挖掘利用中存在的问题和今后的研究方向,希望为高效利用割手密优异抗逆基因资源开展甘蔗多抗逆性聚合育种提供参考。