Induced Homoeologous Pairing for Transfer of Useful Variability for High Grain Fe and Zn from <Emphasis Type="Italic">Aegilops kotschyi</Emphasis> into Wheat

Biofortification of bread wheat by the transfer of useful variability of high grain Fe and Zn from Aegilops kotschyi through induced homoeologous pairing is the most feasible approach to alleviate micronutrient malnutrition worldwide. Deficiency of chromosome 5B in interspecific hybrids allows homoeologous pairing and recombination of chromosomes of wheat with those of the related species. The interspecific hybrid plants without 5B chromosome showed much higher chromosome pairing than did the plants with 5B. The F₁ plants without 5B chromosome were selected and repeatedly backcrossed with wheat cultivar PBW343. The chromosome number of BC₂F₁ plants ranged from 43 to 60 with several univalents and multivalents. Molecular markers and GISH analysis confirmed the introgression of U/S chromosomes of Ae. kotschyi and their fragments in wheat. The BC₂F₂ plants showed up to 125 % increase in Fe and 158 % increase in Zn compared to PBW343 with Lr24 and Yr36. Induced homoeologous pairing in the absence of 5B was found to be an effective approach for transfer of useful variability for enhanced grain Fe and Zn content for biofortification of wheat for high grain micronutrient content.     

Genetic marking of glume color in <Emphasis Type="Italic">Triticum spelta</Emphasis> L. var. <Emphasis Type="Italic">caeruleum</Emphasis> using gliadins

Using gliadins as genetic markers, Triticum spelta L. var. caeruleum accessions were analyzed to identify genetic control of the dark color of glumes. The research material was F₂ and BC₁ plants from crosses between spelt accessions and white-glumed common wheat varieties. The segregation for glume color fitted the monogenic control of the trait. The electrophoretic analysis of gliadins in grains from the hybrid plants has shown that the Gli-Alj* allele in the T. spelta var. caeruleum accessions is linked to the allele for the dark (black) color of glumes at the Rg-A1 locus.     

<Emphasis Type="Italic">Pm62</Emphasis>, an adult-plant powdery mildew resistance gene introgressed from <Emphasis Type="Italic">Dasypyrum villosum</Emphasis> chromosome arm 2VL into wheat

Key message

Pm62, a novel adult-plant resistance (APR) gene against powdery mildew, was transferred from D. villosum into common wheat in the form of Robertsonian translocation T2BS.2VL#5.

Abstract

Powdery mildew, which is caused by the fungus Blumeria graminis f. sp. tritici, is a major disease of wheat resulting in substantial yield and quality losses in many wheat production regions of the world. Introgression of resistance from wild species into common wheat has application for controlling this disease. A Triticum durum-Dasypyrum villosum chromosome 2V#5 disomic addition line, N59B-1 (2n?=?30), improved resistance to powdery mildew at the adult-plant stage, which was attributable to chromosome 2V#5. To transfer this resistance into bread wheat, a total of 298 BC₁F₁ plants derived from the crossing between N59B-1 and Chinese Spring were screened by combined genomic in situ hybridization and fluorescent in situ hybridization, 2V-specific marker analysis, and reaction to powdery mildew to confirm that a dominant adult-plant resistance gene, designated as Pm62, was located on chromosome 2VL#5. Subsequently, the 2VL#5 (2D) disomic substitution line (NAU1825) and the homozygous T2BS.2VL#5 Robertsonian translocation line (NAU1823), with normal plant vigor and full fertility, were identified by molecular and cytogenetic analyses of the BC₁F₂ generation. The effects of the T2BS.2VL#5 recombinant chromosome on agronomic traits were also evaluated in the F₂ segregation population. The results suggest that the translocated chromosome may have no distinct effect on plant height, 1000-kernel weight or flowering period, but a slight effect on spike length and seeds per spike. The translocation line NAU1823 has being utilized as a novel germplasm in breeding for powdery mildew resistance, and the effects of the T2BS.2VL#5 recombinant chromosome on yield-related and flour quality characters will be further assessed.

     

Distribution of the fertility-restoring gene <Emphasis Type="Italic">Rf3</Emphasis> in common and spelt wheat determined by an informative SNP marker

Male sterility induced by the cytoplasm of Triticum timopheevii Zhuk. has shown potential for hybrid seed production in common wheat (Triticum aestivum L.). As hybrids produced by this method are often partially sterile, fertility restoration is crucial for implementing this technology in breeding practice. Several restorer genes were identified, of which Rf3 is one of the most effective genes for achieving restoration. Previous studies located Rf3 on chromosome 1B in common and spelt wheat. However, the distribution of Rf3 in these taxa remained unclear. In the present study, we genetically mapped Rf3 using a BC₁ population derived from CMS-Sperber and the restorer line Primepi (N = 193). After marker validation in four independent BC₁ populations and a diversity panel, we evaluated the distribution of Rf3 in 524 common wheat and 30 European spelt genotypes. In the mapping population, the SNP marker IWB72107 cosegregated with Rf3, whereas IWB14060 was mapped 2.0 cM distal on chromosome 1BS. Surveying the linkage between IWB72107 and Rf3 in the four validation populations revealed map distances that ranged from 0.4 to 2.3 cM. Validation of IWB72107 in the diversity panel showed that it is suitable for marker-assisted selection and related applications. Using this marker, we estimated that 8.8% of the common wheat lines and 66.7% of the spelt cultivars carried the restoring Rf3 allele. We propose that Rf3 explains the restoration capacity of a large proportion of European common wheat lines.     

QTL mapping for tuberous stem formation of kohlrabi (<Emphasis Type="Italic">Brassica oleracea</Emphasis> var. <Emphasis Type="Italic">gongylodes</Emphasis> L.)

The tuberous stem of kohlrabi is an important quantitative trait, which affects its yield and quality. Genetic control of this trait has not yet been unveiled. To identify the QTLs controlling stem swelling of kohlrabi, a BC₁ population of 92 plants was developed from a cross of broccoli DH line GCP04 and kohlrabi var. Seine. A wide range of variation in tuberous stem diameter was observed among the mapping populations. We constructed a genetic map of nine linkage groups (LGs) with different types of markers, spanning a total length of 913.5 cM with an average marker distance of 7.55 cM. Four significant QTLs for radial enlargement of kohlrabi stem, namely, REnBo1, REnBo2, REnBo3, and REnBo4 were detected on C02, C03, C05, and C09, respectively, and accounted for the phenotypic variation of 59% for the stem diameter and 55% for the qualitative grading of tuberous stem in classes. Then, we confirmed the stability of identified QTLs using BC₁S₁ populations derived from the BC₁ plants having heterozygous alleles at the target QTL and homozygous kohlrabi alleles at the remaining QTLs. REnBo1and REnBo2 using 128 plants of BC₁68S₁ and 94 plants of BC₁43S₁, respectively, and REnBo3 and REnBo4 using 152 plants of BC₁57S₁ were detected at the same positions as the respective QTLs of the BC₁ population. Confirmation of QTLs in two successive generations indicates that the QTLs are persistent. The QTLs obtained in this study could be useful in marker-assisted selection of kohlrabi breeding, and to understand the genetic mechanisms of stem swelling and storage organ development in kohlrabi and other Brassica species.     

Salt tolerance conferred by expression of a global regulator IrrE from <Emphasis Type="Italic">Deinococcus radiodurans</Emphasis> in oilseed rape

As salinity is a major threat to sustainable agriculture worldwide, cultivation of salt-tolerant crops becomes increasingly important. IrrE acts as a global regulator and a general switch for stress resistance in Deinococcus radiodurans. In this study, to determine whether the irrE gene can improve the salt tolerance of Brassica napus, we introduced the irrE gene into B. napus by the Agrobacterium tumefaciens-mediated transformation method. Forty-two independent transgenic plants were regenerated. Polymerase chain reaction (PCR) analyses confirmed that the irrE gene had integrated into the plant genome. Northern as well as Western blot analyses revealed that the transgene was expressed at various levels in transgenic plants. Analysis for the T₁ progenies derived from four independent transformants showed that irrE had enhanced the salt tolerance of T₁ in the presence of 350 mM NaCl. Furthermore, under salt stress, transgenic plants accumulated more compatible solutes (proline) and a lower level of malondialdehyde (MDA), and they had higher activities of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). However, agronomic traits were not affected by irrE gene overexpression in the transgenic B. napus plants. This study indicates that the irrE gene can improve the salt tolerance of B. napus and represents a promising candidate for the development of crops with enhanced salt tolerance by genetic engineering.     

Fine mapping of QTL <Emphasis Type="Italic">qCTB10</Emphasis>-<Emphasis Type="Italic">2</Emphasis> that confers cold tolerance at the booting stage in rice

Key message

The QTL qCTB10 - 2 controlling cold tolerance at the booting stage in rice was delimited to a 132.5 kb region containing 17 candidate genes and 4 genes were cold-inducible.

Abstract

Low temperature at the booting stage is a major abiotic stress-limiting rice production. Although some QTL for cold tolerance in rice have been reported, fine mapping of those QTL effective at the booting stage is few. Here, the near-isogenic line ZL31-2, selected from a BC₇F₂ population derived from a cross between cold-tolerant variety Kunmingxiaobaigu (KMXBG) and the cold-sensitive variety Towada, was used to map a QTL on chromosome 10 for cold tolerance at the booting stage. Using BC₇F₃ and BC₇F₄ populations, we firstly confirmed qCTB10-2 and gained confidence that it could be fine mapped. QTL qCTB10-2 explained 13.9 and 15.9% of the phenotypic variances in those two generations, respectively. Using homozygous recombinants screened from larger BC₇F₄ and BC₇F₅ populations, qCTB10-2 was delimited to a 132.5 kb region between markers RM25121 and MM0568. 17 putative predicted genes were located in the region and only 5 were predicted to encode expressed proteins. Expression patterns of these five genes demonstrated that, except for constant expression of LOC_Os10g11820, LOC_Os10g11730, LOC_Os10g11770, and LOC_Os10g11810 were highly induced by cold stress in ZL31-2 compared to Towada, while LOC_Os10g11750 showed little difference. Our results provide a basis for identifying the genes underlying qCTB10-2 and indicate that markers linked to the qCTB10-2 locus can be used to improve the cold tolerance of rice at the booting stage by marker-assisted selection.

     

Enhanced salinity stress tolerance in transgenic chilli pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) plants overexpressing the wheat antiporter (<Emphasis Type="Italic">TaNHX2</Emphasis>) gene

Transgenic chilli pepper (Capsicum annuum L.) plants tolerant to salinity stress were produced by introducing the wheat Na⁺/H⁺ antiporter gene (TaNHX2) via Agrobacterium-mediated transformation. Cotyledonary explants were infected with Agrobacterium tumefaciens strain LBA4404 harboring a binary vector pBin438 that contains a wheat antiporter (TaNHX2) gene driven by the double CaMV 35S promoter and NPT II gene as a selectable marker. PCR and semiquantitative RT-PCR analysis confirmed that the TaNHX2 gene had been integrated and expressed in the T₁ generation of transgenic pepper plants as compared to the non-transformed plants. Southern blot analysis further verified the integration and presence of TaNHX2 gene in the genome of chilli pepper plants. Biochemical assays of these transgenic plants revealed enhanced levels of proline, chlorophyll, superoxide dismutase, ascorbate peroxidase, relative water content, and reduced levels of hydrogen peroxide (H₂O₂), malondialdehyde compared to wild-type plants under salt stress conditions. The present investigation clearly showed that overexpression of the TaNHX2 gene enhanced salt stress tolerance in transgenic chilli pepper plants.     

Improved versions of rice maintainer line,APMS 6B,possessing two resistance genes, <Emphasis Type="Italic">Xa21</Emphasis> and <Emphasis Type="Italic">Xa38</Emphasis>, exhibit high level of resistance to bacterial blight disease

APMS 6B is the stable maintainer of the CMS line APMS 6A, which is the female parent of the popular Indian rice hybrid DRRH 3. APMS 6B has good combining ability and plant stature but is highly susceptible to bacterial blight (BB) disease. In order to improve the BB resistance of APMS 6B, we pyramided two major, dominant BB resistance genes, Xa21 and Xa38, through marker-assisted backcross breeding (MABB). Improved Samba Mahsuri (ISM) was used as the donor for Xa21 while PR 114 (Xa38) served as the donor for Xa38. Individual crosses [APMS 6B/ISM and APMS 6B/PR 114 (Xa38)] were performed, and true F₁ plants were then backcrossed with APMS 6B and the MABB process was continued till BC₃. A single positive BC₃F₁ plant identified from both the crosses with maximum genotypic and phenotypic similarity with APMS 6B was selfed to generate BC₃F₂s. At BC₃F₂ generation, plants homozygous for either Xa21 or Xa38 were identified and further confirmed for the absence of two major fertility restorer genes, Rf3 and Rf4. A single such homozygous BC₃F₂ plant, each from both the crosses, was then inter-mated to generate ICF₁s (inter-cross F₁s). Selected ICF₁ plants possessing both the BB resistance genes were selfed to generate ICF₂s. A total of 42 ICF₂ plants homozygous for both Xa21 and Xa38 were identified and screened with parental polymorphic SSR markers to identify the best F₂ plants having the maximum recurrent parent genome recovery. Twelve best ICF₂ plants were advanced up to ICF₅. The ICF₅ lines displayed very high level of BB resistance and were similar to APMS 6B in terms of agro-morphological characters. Further, most of these lines also showed complete maintenance ability and such lines are being advanced for conversion to WA-CMS lines.     

Wheat methionine sulfoxide reductase genes and their response to abiotic stress

The wheat cultivar Shanrong no. 3 (cv. SR3) tolerates both salinity and drought stress more effectively than does its progenitor cultivar Jinan 177 (cv. JN177). When the cultivars are subjected to stress, a number of genes encoding methionine sulfoxide reductase (MSRs) are known to be upregulated in SR3. Here, a set of 12 full length Triticum aestivum MSR (TaMSR) cDNAs have been isolated from cv. SR3. The genes were transcribed in the wheat root, stem, and leaf in plants sampled at various developmental stages. Those induced by salinity and drought harbored known stress-responsive cis elements in their promoter region. The constitutive expression in Arabidopsis thaliana of four MSRs which were induced by salt and drought in microarray assay showed that the product of one (TaMSRA2) heightened the plant’s tolerance to NaCl, methylviologen (MV), and abscisic acid, that of the second (TaMSRA5) enhanced salinity tolerance, that of the third (TaMSRB1.1) increased tolerance to salinity, MV and H₂O₂, and that of the fourth (TaMSRB5.1) increased tolerance to both salinity and mannitol. The effect of the presence in A. thaliana of TaMSRB1.1 was to suppress the accumulation of reactive oxygen species and to increase the intracellular content of soluble sugars.     

Development of a robust marker for <Emphasis Type="Italic">Psy-1</Emphasis> homoeologs and its application in improvement of yellow pigment content in durum wheat

Phytoene synthase-1 (Psy-1) homoeologs are associated with yellow pigment content (YPC) in endosperm of durum and bread wheat. In the present study, microsatellite variation in promoter region of Psy-A1 was identified in durum wheat and marker Psy-1SSR, targeting the microsatellite variation was developed which amplifies variation in Psy-A1 and Psy-B1 loci simultaneously. Psy-A1SSR was mapped within QYp.macs-7A, a major QTL for YPC identified earlier in PDW 233/Bhalegaon 4 population. Marker Psy-A1SSR was further validated in two different RIL populations and a set of 222 tetraploid wheat accessions including less cultivated tetraploid wheat species. Eight alleles of Psy-A1SSR were identified in 222 wheat accessions, while seven alleles were observed for Psy-B1SSR. Variation at Psy-A1SSR showed significant association with YPC, whereas no association was observed with Psy-B1SSR. Marker-assisted introgression of Psy-A1SSRe allele from PDW 233, to durum wheat cultivars MACS 3125 and HI 8498 resulted in improvement of YPC. Backcrossed BC₃F_2:4 and BC₂F_2:3 lines selected using Psy-A1SSR showed 89 to 98% gain in YPC over recurrent parents indicating robustness of marker. The marker can thus be utilized in marker-assisted improvement of YPC in durum wheat cultivars.     

Identification of <Emphasis Type="Italic">Pm58</Emphasis> from <Emphasis Type="Italic">Aegilops tauschii</Emphasis>

Key message

A novel powdery mildew-resistance gene, designated Pm58, was introgressed directly from Aegilops tauschii to hexaploid wheat, mapped to chromosome 2DS, and confirmed to be effective under field conditions. Selectable KASP? markers were developed for MAS.

Abstract

Powdery mildew caused by Blumeria graminis (DC.) f. sp. tritici (Bgt) remains a significant threat to wheat (Triticum aestivum L.) production. The rapid breakdown of race-specific resistance to Bgt reinforces the need to identify novel sources of resistance. The d-genome species, Aegilops tauschii, is an excellent source of disease resistance that is transferrable to T. aestivum. The powdery mildew-resistant Ae. tauschii accession TA1662 (2n?=?2x?=?DD) was crossed directly with the susceptible hard white wheat line KS05HW14 (2n?=?6x?=?AABBDD) followed by backcrossing to develop a population of 96 BC₂F₄ introgression lines (ILs). Genotyping-by-sequencing was used to develop a genome-wide genetic map that was anchored to the Ae. tauschii reference genome. A detached-leaf Bgt assay was used to screen BC₂F_4:6 ILs, and resistance was found to segregate as a single locus (χ?=?2.0, P value?=?0.157). The resistance gene, referred to as Pm58, mapped to chromosome 2DS. Pm58 was evaluated under field conditions in replicated trials in 2015 and 2016. In both years, a single QTL spanning the Pm58 locus was identified that reduced powdery mildew severity and explained 21% of field variation (P value?<?0.01). KASP? assays were developed from closely linked GBS-SNP markers, a refined genetic map was developed, and four markers that cosegregate with Pm58 were identified. This novel source of powdery mildew-resistance and closely linked genetic markers will support efforts to develop wheat varieties with powdery mildew resistance.

     

Breeding lines of the Indian mega-rice variety,MTU 1010, possessing protein kinase <Emphasis Type="Italic">OsPSTOL</Emphasis> (<Emphasis Type="Italic">Pup1</Emphasis>), show better root system architecture and higher yield in soils with low phosphorus

MTU 1010 is a high-yielding mega-variety of rice grown extensively in India. However, it does not perform well in soils with low phosphorus (P) levels. With an objective to improve MTU 1010 for tolerance to low soil P, we have transferred Pup1, a major quantitative trait locus (QTL) associated with tolerance from another mega-variety, Swarna, through marker-assisted backcross breeding (MABB). Foreground selection of the F₁ and backcross plants was performed with the co-dominant, closely linked CAPS marker, K20-2, while two flanking markers RM28011 and RM28157 were utilized for recombinant selection. At each backcross generation, positive plants were also analyzed with a set of 85 parental polymorphic SSR markers to identify the QTL-positive plants possessing maximum introgression of MTU 1010 genome. At BC₂F₁, the best backcross plant was selfed to generate BC₂F₂s. Among them, the plants homozygous for Pup1 (n?=?22) were reconfirmed using the functional marker for Pup1, viz., K46-1, and they were advanced through pedigree method of selection until BC₂F₆ generation. A total of five elite BC₂F₆ lines, possessing Pup1 and phenotypically similar to MTU 1010, were screened in the low soil P plot and normal plot (with optimum soil P levels) during wet season, 2016. All the selected lines showed better performance under low P soil with more number of productive tillers, better root system architecture, and significantly higher yield (>?390%) as compared to MTU 1010. Further, under normal soil, the lines were observed to be similar to or better than MTU 1010 for most of the agro-morphological traits and yield. This study represents the successful application of marker-assisted selection for improvement of tolerance to low soil P in a high-yielding Indian rice variety.     

Marker-assisted introgression of the major bacterial blight resistance gene, <Emphasis Type="Italic">Xa21</Emphasis> and blast resistance gene, <Emphasis Type="Italic">Pi54</Emphasis> into RPHR-1005, the restorer line of the popular rice hybrid,DRRH3

The elite Indian rice hybrid, DRRH3 is highly susceptible to two major diseases, bacterial blight (BB) and blast, which limit its productivity significantly. In the present study, we have introgressed two major genes, viz., Xa21 and Pi54 conferring resistance against BB and blast, respectively into RPHR-1005, the male parent of DRRH3 through marker-assisted backcross breeding (MABB) and analyzed the backcross derived plants for their resistance against BB and blast. RPBio Patho-2 was used as a donor for both the resistance genes. Gene-specific markers were used for the foreground selection of Xa21 and Pi54 at each stage of backcrossing and markers specific for the major fertility restorer genes, Rf3 and Rf4 were used only at BC₁F₁ generation for foreground selection. Background selection was done using 62 polymorphic SSR markers and marker-assisted backcrossing was continued till BC₃ generation. At BC₃F₄, through intensive phenotype-based selections 15 promising lines (ABLs) possessing high level of resistance against BB and blast, high yield, fine-grain type, complete fertility restoration along with better panicle exsertion and taller plant type as compared to RPHR-1005 were identified and test crossed with APMS 6 A, the female parent of DRRH3. The newly derived hybrids (i.e. improved versions of DRRH3) were observed to possess high level of resistance against BB and blast along with medium-slender grain type and yield level better than or equivalent to that of DRRH3. Our study exemplifies the utility of MABB for targeted improvement of multiple traits in hybrid rice.     

Physical mapping of DNA markers linked to stem rust resistance gene <Emphasis Type="Italic">Sr47</Emphasis> in durum wheat

Key message

Markers linked to stem rust resistance gene Sr47 were physically mapped in three small Aegilops speltoides chromosomal bins. Five markers, including two PCR-based SNP markers, were validated for marker-assisted selection.

Abstract

In durum wheat (Triticum turgidum subsp. durum), the gene Sr47 derived from Aegilops speltoides conditions resistance to race TTKSK (Ug99) of the stem rust pathogen (Puccinia graminis f. sp. tritici). Sr47 is carried on small interstitial translocation chromosomes (Ti2BL-2SL-2BL·2BS) in which the Ae. speltoides chromosome 2S segments are divided into four bins in genetic stocks RWG35, RWG36, and RWG37. Our objective was to physically map molecular markers to bins and to determine if any of the molecular markers would be useful in marker-assisted selection (MAS). Durum cultivar Joppa was used as the recurrent parent to produce three BC₂F₂ populations. Each BC₂F₂ plant was genotyped with markers to detect the segment carrying Sr47, and stem rust testing of BC₂F₃ progeny with race TTKSK confirmed the genotyping. Forty-nine markers from published sources, four new SSR markers, and five new STARP (semi-thermal asymmetric reverse PCR) markers, were evaluated in BC₂F₂ populations for assignment of markers to bins. Sr47 was mapped to bin 3 along with 13 markers. No markers were assigned to bin 1; however, 7 and 13 markers were assigned to bins 2 and 4, respectively. Markers Xrwgs38a, Xmag1729, Xwmc41, Xtnac3119, Xrwgsnp1, and Xrwgsnp4 were found to be useful for MAS of Sr47. However, STARP markers Xrwgsnp1 and Xrwgsnp4 can be used in gel-free systems, and are the preferred markers for high-throughput MAS. The physical mapping data from this study will also be useful for pyramiding Sr47 with other Sr genes on chromosome 2B.

     

Strains of <Emphasis Type="Italic">Bacillus</Emphasis> ssp. regulate wheat resistance to <Emphasis Type="Italic">Septoria nodorum</Emphasis> Berk.

The ability of Bacillus subtilis Cohn and Bacillus thuringiensis Berliner to induce systemic resistance in wheat plants to the casual agent of Septoria nodorum Berk., blotch has been studied. It has been shown that strains of Bacillus ssp. that possess the capacity for endophytic survival have antagonistic activity against this pathogen in vitro. A reduction of the degree of Septoria nodorum blotch development on wheat leaves under the influence of Bacillus spp. was accompanied by the suppression of catalase activity, an increase in peroxidase activity and H₂O₂ content, and expression of defence related genes such us PR-1, PR-6, and PR-9. It has been shown that B. subtilis 26 D induces expression levels of wheat pathogenesis-related (PR) genes which marks a SA-dependent pathway of sustainable development and that B. thuringiensis V-5689 and V-6066 induces a JA/ET-dependent pathway. These results suggest that these strain Bacillus spp. promotes the formation of wheat plant resistance to S. nodorum through systemic activation of the plant defense system. The designed bacterial consortium formed a complex biological response in wheat plants infected phytopathogen.     

Transfer and mapping of the heat tolerance component traits of <Emphasis Type="Italic">Aegilops speltoides</Emphasis> in tetraploid wheat <Emphasis Type="Italic">Triticum durum</Emphasis>

Aegilops speltoides is an important genetic resource for wheat improvement and has high levels of heat tolerance. A heat-tolerant accession of Ae. speltoides pau3809 was crossed with Triticum durum cv. PDW274, and BC₂F_4-6 backcross introgression lines (BILs) were developed, phenotyped for important physiological traits, genotyped using SSR markers and used for mapping the QTL governing heat tolerance component traits. A set of 90 BILs was selected from preliminary evaluation of a broader set of 262 BILs under heat stress. Phenotyping was conducted for physiological traits such as cell membrane thermostability, chlorophyll content, acquired thermotolerance, canopy temperature and stay green. Much variation for these traits was observed in random as well as selected sets of BILs, and comparison of the BILs with the recurrent parent showed improvement for these traits under normal as well as heat stress conditions, indicating that introgressions from Ae. speltoides might have led to the improvement in the heat tolerance potential of the BILs. Introgression profiling of the 90 BILs using SSR markers identified Ae. speltoides introgression on all the 14 chromosomes with introgressions observed on A as well as B genome chromosomes. QTL mapping identified loci for various heat tolerance component traits on chromosomes 2B, 3A, 3B, 5A, 5B and 7A at significant LOD scores and with phenotypic contributions varying from 11.1 to 28.7 % for different traits. The heat-tolerant BILs and QTL reported in the present study form a potential resource that can be used for wheat germplasm enhancement for heat stress tolerance.     

Tolerance to soil water stress by <Emphasis Type="Italic">Oryza sativa</Emphasis> cv. IR20 was improved by expression of <Emphasis Type="Italic">Wsi18</Emphasis> gene locus from <Emphasis Type="Italic">Oryza nivara</Emphasis>

Wild rice genotypes are rich in genetic diversity. This has potential to improve agronomic rice by allele mining for superior traits. Late embryogenesis abundant (LEA) proteins are often associated with desiccation tolerance and stress signalling. In the present study, a group 3 LEA gene, Wsi18 from the wild rice Oryza nivara was expressed under its own inducible promoter element in stress susceptible cultivated indica rice (cv. IR20). The resulting transgenic plants cultivated in a greenhouse showed enhanced tolerance to soil water deficit. Transgenic plants had higher grain yield, plant survival rate, and shoot relative water content compared to wild type (WT) IR20. Cell membrane stability index, proline and soluble sugar content were also greater in transgenic than WT plants under water stress. These results demonstrate the potential for improving SWS tolerance in agronomically important rice cultivar by incorporating Wsi18 gene from a wild rice O. nivara.     

Identification and mapping of <Emphasis Type="Italic">MLIW30</Emphasis>, a novel powdery mildew resistance gene derived from wild emmer wheat

Powdery mildew, caused by Blumeria graminis f.sp. tritici (Bgt), is a destructive foliar disease of common wheat in areas with cool or maritime climates. Wild emmer wheat, Triticum turgidum ssp. dicoccoides, the progenitor of both domesticated tetraploid durum wheat and hexaploid bread wheat, harbors abundant genetic diversity related to resistance to powdery mildew that can be utilized for wheat improvement. An F₂ segregating population was obtained from a cross between resistant bread wheat line 2L6 and susceptible cultivar Liaochun 10, after which genetic analysis of F₂ and F₂-derived F₃ families was performed by inoculating plants with isolate Bgt E09. The results of this experiment demonstrated that powdery mildew resistance in 2L6, which was derived from wild emmer wheat accession IW30, was controlled by a single dominant gene, temporarily designated MLIW30. Nineteen SSR markers and two STS markers linked with MLIW30 were acquired by applying bulked segregant analysis. Finally, MLIW30 was located to the long arm of chromosome 4A and found to be flanked by simple sequence repeat markers XB1g2000.2 and XB1g2020.2 at 0.1 cM. Because no powdery mildew resistance gene in or derived from wild emmer wheat has been reported in wheat chromosome 4A, MLIW30 might be a novel Pm gene.