Lili Xu

Genetic diversity of HMW glutenin subunits in diploid, tetraploid and hexaploid Triticum species.

The genetic variations of high-molecular-weight (HMW) glutenin subunits in 1051 accessions of 13 Triticum subspecies were investigated using sodium dodecyl sulfate polyacrylamide-gel electrophoresis. A total of 37 alleles were detected, resulting in 117 different allele combinations, among which 20, 68 and 29 combinations were observed in diploid, tetraploid and hexaploid wheats, respectively. Abundance and frequency of allele and combinations in tetraploid wheats were higher than these in hexaploid wheats. Allele Glu-A1c was the most frequent subunit at Glu-A1 locus in tetraploid and hexaploid wheats. Consequently, the results also suggested that the higher variations occurred at Glu-B1 locus compared to Glu-A1 and Glu-D1. Therefore, carthlicum wheat possessing the allele 1Ay could be presumed a special evolutional approach distinguished from other tetraploid species. Furthermore, this provides a convenient approach of induction of the 1Ay to common wheat through direct cross with carthlicum wheat. Alleles Glu-B1c and Glu-B1i generally absent in tetraploid wheats were also found in tetraploid wheats. Our results implied that tetraploid and hexaploid wheats were distinguished in dendrogram, whereas carthlicum and spelta wheats and however displayed the unique performance. In addition, founder effect, no-randomness of diploidization, mutation and artificial selection could cause allele distribution of HMW-GS in Triticum. All alleles of HMW-GS in Triticum could be further utilized through hybrid in the quality improvement of common wheat.

Molecular cytogenetic characterization and stripe rust response of a trigeneric hybrid involving Triticum, Psathyrostachys, and Thinopyrum

Trigeneric hybrids offer opportunities to transfer alien traits into cultivated wheat. In this study, a new trigeneric hybrid involving species of Triticum, Psathyrostachys, and Thinopyrum was synthesized by crossing Triticum aestivum L. (wheat)--Thinopyrum intermedium (Host) Barkworth & D.R. Dewey amphiploid Zhong 3 with wheat--Psathyrostachys huashanica Keng ex Kuo amphiploid PHW-SA. Crossability of the two amphiploids was 19.74%, and the fertility of the hybrid was 16.20%. The mean meiotic configuration of the trigeneric hybrid (2n=56) was 13.06 I+17.24 IIring+3.73 IIrod+0.28 III+0.04 IV. GISH analysis indicated that the trigeneric F1 had seven P. huashanica chromosomes and seven Th. intermedium chromosomes. The mean chromosome numbers of F2, F3, and F4 progenies were 2n=49.24, 2n=48.13, and 2n=46.78, respectively, a gradual decrease. GISH analysis revealed that most F2 and F3 plants had 2–10 Th. intermedium chromosomes and 0–4 P. huashanica chromosomes. In the F4 progenies, 1–7 Th. intermedium chromosomes were labeled, but no P. huashanica chromosomes were detected. It seems that Th. intermedium chromosomes are more likely than P. huashanica chromosomes to be transmitted to the progenies. The stripe rust response of PHW-SA was expressed in the F1 and some F2 and F3 progenies. The trigeneric hybrid could be a useful bridge for transfering P. huashanica and Th. intermedium chromosomes to common wheat.

Divergent Development of Hexaploid Triticale by a Wheat – Rye –Psathyrostachys huashanica Trigeneric Hybrid Method

Hexaploid triticale is an important forage crop and a promising energy plant. Some forms were previously reported for developing the hexaploid triticale, such as crossing tetraploid wheat or hexaploid wheat with rye, crossing hexaploid triticale and/or hexaploid wheat with octoploid triticale, and spontaneously appearing in the selfed progenies of octoploid triticale. In the present study, we developed an effective method for production of diverse types of hexaploid triticale via wheat—rye—Psathyrostachys huashanica trigeneric hybrid. Genomic in situ hybridization (GISH) and fluorescence in situ hybridization (FISH) karyotyping revealed that D genome chromosomes were completely eliminated and the whole A, B, and R genome chromosomes were retained in three lines. More interestingly, the composite genome of the line K14-489-2 consisted of complete A and B genomes and chromosomes 1D, 2R, 3R, 4R, 5R, 6R, and 7R, that of line K14-491-2 was 12 A-genome (1A-6A), 14 B-genome (1B-7B), 12 R-genome (1R-3R, 5R-7R), and chromosomes 1D and 3D, and that of the line K14-547-1 had 26A/B and 14R chromosomes, plus one pair of centric 6BL/2DS translocations. This finding implies that some of D genome chromosomes can be spontaneously and stably incorporated into the hexaploid triticale. Additionally, a variety of high-molecular-weight glutenin subunits (HMW-GS) compositions were detected in the six hexaploid triticale lines, respectively. Besides, compared with its recurrent triticale parent Zhongsi828, these lines showed high level of resistance to stripe rust (Puccinia striiformis f. sp. tritici, Pst) pathogens prevalent in China, including V26/Gui 22. These new hexaploid triticales not only enhanced diversification of triticale but also could be utilized as valuable germplasm for wheat improvement.

Characterization of wheat-Psathyrostachys huashanica small segment translocation line with enhanced kernels per spike and stripe rust resistance

Psathyrostachys huashanica Keng (2n = 2x = 14, NsNs), a distant wild relative of common wheat, possesses rich potentially valuable traits, such as disease resistance and more spikelets and kernels per spike, that could be useful for wheat genetic improvement. Development of wheat - P. huashanica translocation lines will facilitate its practical utilization in wheat breeding. In the present study, a wheat - P. huashanica small segmental translocation line, K-13-835-3, was isolated and characterized from the BC1F5 population of a cross between wheat - P. huashanica amphiploid PHW-SA and wheat cultivar CN16. Cytological studies showed that the mean chromosome configuration of K-13-835-3 at meiosis was 2n = 42 = 0.10 I + 19.43 II (ring) + 1.52 II (rod). GISH analyses indicated that chromosome composition of K-13-835-3 included 40 wheat chromosomes and a pair of wheat - P. huashanica translocation chromosomes. FISH results demonstrated that the small segment from an unidentified P. huashanica chromosome was translocated into wheat chromosome arm 5DS, proximal to the centromere region of 5DS. Compared with the cultivar wheat parent CN16, K-13-835-3 was highly resistant to stripe rust pathogens prevalent in China. Furthermore, spikelets and kernels per spike in K-13-835-3 were significantly higher than those of CN16 in two growing seasons. These results suggest that the desirable genes from P. huashanica were successfully transferred into CN16 background. This translocation line could be used as novel germplasm for high-yield and, eventually, resistant cultivar breeding.

Evaluation of geographic distribution of high-molecular-weight glutenin subunits (HMW-GS) in wild and cultivated Triticum species

Variation of high-molecular-weight glutenin subunit (HMW-GS) in 632 wild and cultivated Triticum accessions was investigated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. A total of 11 alleles of HMW-GS in diploid species, 22 in tetraploid species, and 15 in hexaploid species were detected. Diploid species on Glu-1A locus and tetraploid species Glu-1B locus showed the highest diversity, respectively. Tetraploid species had the highest level of diversity on three Glu-1 loci, followed by hexaploid and diploid, based on Shannon’s information index, Nei’s genetic diversity, and percentage of polymorphic loci. Molecular variance analysis confirmed main variance of HMW-GS within species, regions, and locations, respectively. Variance among species and regions was enhanced gradually with the increase of ploidy. Significant non-random distributions between the phylogenic trees of HMW-GS and the locations of accessions were tested by GenGIS software, indicated that geographic factors played an important role along the different orientations in the spread of Triticum species. We found one original diversified center in diploid what located around Elazig, Malatya, Gaziantep, Urfa, and Kiziltepe in Turkey, and three diversified centers in tetraploid wheat, including Turkey–Armenia–Georgia–Iran, Portugal–Spain, and Ethiopia, respectively, and two diversified adjacent areas between Turkey and Switzerland and around Turkey, Georgia, and Armenia. The original center of diploid species located in southeast Turkey, where the unexpressed 1Ay subunit was mainly distributed in T. urartu, could be one of the candidate regions of polyploidization of Triticum L. The regional distribution of HMW-GS and species also provided geographic evidences for the existence of founder effect on the spread of Triticum species. The present study suggests that integrating genetic diversity with geographic characterization in Triticum could very useful for collection, conservation, and utilization, as well as for research microevolution and domestication.

FISH-Based Markers Enable Identification of Chromosomes Derived From Tetraploid Thinopyrum elongatum in Hybrid Lines

Tetraploid Thinopyrum elongatum, which has superior abiotic stress tolerance characteristics, and exhibits resistance to stripe rust, powdery mildew, and Fusarium head blight, is a wild relative of wheat and a promising source of novel genes for wheat improvement. Currently, a high-resolution Fluorescence in situ hybridization (FISH) karyotype of tetraploid Th. elongatum is not available. To develop chromosome-specific FISH-based markers, the hexaploid Trititrigia 8801 and two accessions of tetraploid Th. elongatum were characterized by different repetitive sequences probes. We found that all E-genome chromosomes could be unambiguously identified using a combination of pSc119.2, pTa535, pTa71, and pTa713 repeats, and the E-genome chromosomes of the wild accessions and the partial amphiploid failed to exhibit any significant variation in the probe hybridization patterns. To verify the validation of these markers, the chromosome constitution of eight wheat- Th. elongatum hybrid derivatives were analyzed. We revealed that these probes could quickly detect wheat and tetraploid Th. elongatum chromosomes in hybrid lines. K16-712-1-2 was a 1E (1D) chromosome substitution line, K16-681-4 was a 2E disomic chromosome addition line, K16-562-3 was a 3E, 4E (3D, 4D) chromosome substitution line, K15-1033-8-2 contained one 4E, two 5E, and one 4ES⋅1DL Robertsonian translocation chromosome, and four other lines carried monosomic 4E, 5E, 6E, and 7E chromosome, respectively. Furthermore, the E-genome specific molecular markers analysis corresponded perfectly with the FISH results. The developed FISH markers will facilitate rapid identification of tetraploid Th. elongatum chromosomes in wheat improvement programs and allow appropriate alien chromosome transfer.

Cytogenetic study and stripe rust response of the derivatives from a wheat – Thinopyrum intermedium – Psathyrostachys huashanica trigeneric hybrid

To transfer multiple desirable alien genes into common wheat, we previously reported a new trigeneric hybrid synthesized by crossing a wheat - Thinopyrum intermedium partial amphiploid with wheat - Psathyrostachys huashanica amphiploid. Here, the meiotic behavior, chromosome constitution, and stripe rust resistance of F5 derivatives from the wheat - Th. intermedium - P. huashanica trigeneric hybrid were studied. Cytological analysis indicated the F5 progenies had chromosome numbers of 42-50 (average 44.96). The mean meiotic configuration was 1.28 univalents, 21.74 bivalents, 0.04 trivalents, and 0.02 tetravalents per pollen mother cell. In 2n = 42 lines, the average pairing configuration was 0.05 I + 19.91 II (ring) + 1.06 II (rod) + 0.003 IV, suggesting these lines were cytologically stable. Most lines with 2n = 43, 44, 46, 48, or 50, bearing a high frequency of univalents or multivalents, showed abnormal meiotic behavior. Genomic in situ hybridization karyotyping results revealed that 25 lines contained 1-7 Th. intermedium chromosomes, but no P. huashanica chromosomes were found among the 27 self-pollinated progenies. At meiosis, univalents (1-5) possessing Th. intermedium hybridization signals were detected in 19 lines. Bivalents (1-3) expressing fluorescence signals were observed in 12 lines. Importantly, 21 lines harbored wheat - Th. intermedium chromosomal translocations with various alien translocation types. Additionally, two homozygous lines, K13-668-10 and K13-682-12, possessed a pair of wheat - Th. intermedium small fragmental translocations. Compared with the recurrent parent Zhong 3, most lines showed high resistance to the stripe rust (Puccinia striiformis f. sp. tritici) pathogens prevalent in China, including race V26/Gui22. This paper reports a highly efficient technical method for inducing alien translocation between wheat and Th. intermedium by trigeneric hybridization. These lines might be potentially valuable germplasm resources for further wheat improvement.

Cytogenetic Behavior of Trigeneric Hybrid Progeny Involving Wheat, Rye and Psathyrostachys huashanica

Trigeneric hybrids are commonly used as bridges to transfer genes from some wild species to cultivated wheat and to measure the genomic interaction between donor species. We previously reported that trigeneric germplasms were produced by crossing wheat-Psathyrostachys huashanica amphiploids (PHW-SA, 2n = 8x = 56, AABBDDNsNs) with hexaploid triticale (Zhongsi 828, 2n = 6x = 42, AABBRR). In the present study, chromosome pairing behavior and the genome constitution of the F4 progenies of wheat-rye-P. huashanica trigeneric hybrids were studied. Cytological analysis showed that the chromosome number of F4 progenies ranged from 39 to 46, and 57.5% of them had 42 chromosomes. The mean meiotic configuration of F4 lines was 1.71 univalents, 20.26 bivalents, 0.04 trivalents, and 0.001 quadrivalents per pollen mother cell. Among the lines with 2n = 42, the average pairing configuration was 1.21 univalents, 16.22 ring bivalents, 4.16 rod bivalents, and 0.01 trivalents. This result indicated that these lines were cytologically stable. Other lines with 2n = 39, 40, 41, 43, 44, 45, and 46, bearing a high number of univalents or multivalents, showed abnormal meiotic behavior. Genomic in situ hybridization (GISH) revealed that all F4 lines had 11-14 rye chromosomes, but no P. huashanica chromosomes. The complete set of 14 rye chromosomes was found in 19 lines. At meiosis, GISH detected 1-6 univalents with hybridization signals of rye in 13 lines. Bivalents with fluorescence signals were identified in each line, ranging from 3 to 7. A quadrivalent with hybridization signals was observed in only 1 line, K13-714-8. Lagging chromosomes, chromosome bridges, micronuclei, and chromosome fragments hybridizing with the probe were not discovered in any of the lines. These results inferred that the behavior of rye chromosomes was normal during meiosis. In addition, 21 lines of 2n = 42 (91.3%) with 12 or 14 rye chromosomes, always contained 6 or 7 bivalents bearing fluorescence signals. This suggested that the rye chromosomes exhibiting complete pairing in these lines were cytologically stable during meiosis and may therefore be considered as new hexaploid triticales. Thus, these lines might be potential materials for further hexaploid triticale improvement.

Evolutionary History of Triticum petropavlovskyi Udacz. et Migusch. Inferred from the Sequences of the 3-Phosphoglycerate Kinase Gene

Single- and low-copy genes are less likely to be subject to concerted evolution. Thus, they are appropriate tools to study the origin and evolution of polyploidy plant taxa. The plastid 3-phosphoglycerate kinase gene (Pgk-1) sequences from 44 accessions of Triticum and Aegilops, representing diploid, tetraploid, and hexaploid wheats, were used to estimate the origin of Triticum petropavlovskyi. Our phylogenetic analysis was carried out on exon+intron, exon and intron sequences, using maximum likelihood, Bayesian inference and haplotype networking. We found the D genome sequences of Pgk-1 genes from T. petropavlovskyi are similar to the D genome orthologs in T. aestivum, while their relationship with Ae. tauschii is more distant. The A genome sequences of T. petropavlovskyi group with those of T. polonicum, but its Pgk-1 B genome sequences to some extent diverge from those of other species of Triticum. Our data do not support for the origin of T. petropavlovskyi either as an independent allopolyploidization event between Ae. tauschii and T. polonicum, or as a monomendelian mutation in T. aestivum. We suggest that T. petropavlovskyi originated via spontaneous introgression from T. polonicum into T. aestivum. The dating of this introgression indicates an age of 0.78 million years; a further mutation event concerning the B genome occurred 0.69 million years ago.

Cytogenetic identification of wheat-Psathyrostachys huashanica amphiploid × triticale progenies for English grain aphid resistance

English grain aphid (EGA, Sitobion avenae Fabricius) is an important pest in wheat (Triticum aestivum L.). To develop EGA-resistant varieties, introducing the desirable genes from related species is regarded as an efficient avenue. In this study, the F1, F2 and F3 plants derived from the cross of EGA-susceptible wheat-Psathyrostachys huashanica Keng ex Kuo amphiploid (PHW-SA, AABBDDNsNs) and EGA-resistant triticale (Zhongsi 828, AABBRR) were analyzed for EGA resistance. Consequently, PHW-SA was moderately susceptible while Zhongsi 828 and their F1 hybrids were immune, suggesting that the resistance is dominant. All the F2 plants showed high resistance or immunity over two years, indicating that EGA resistance genes are more likely carried by the rye (Secale cereale L.) chromosomes rather than the genomes A or B of Zhongsi 828. In the F3 generation, 25 of 239 lines became susceptible. Giemsa C-banding patterns revealed that these F3 lines had 38-40 chromosomes, including complete rye genome except 5R (and 2R in five lines). Genomic in situ hybridization analysis confirmed this result. During meiosis, all the chromosomes formed bivalents. Six bivalents in 20 lines and five bivalents in five lines were characterized from rye. In contrast, their F2 parental lines had 42 chromosomes (21 bivalents), containing 1R-7R of rye. No P. huashanica chromosomes were detected. Therefore, we propose that the rye chromosome 5R may be related to EGA resistance.