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Although significant progress has been made on Agrobacterium -mediated wheat transformation, the current methodologies use immature embryos as recipient tissues, a process which is labor intensive, time consuming and expensive. In this study, we have managed to develop an Agrobacterium -based transformation scheme using explants derived from mature embryos. Based on transient expression of β -glucuronidase (GUS) marker, mature embryo halves prepared from freshly imbibed seeds were generally most susceptible to Agrobacterium -mediated T-DNA transfer. According to the results of callus induction and shoot production, Yumai 66 and Lunxuan 208 showed higher selection and regeneration efficiency than Bobwhite. In line with this finding, fertile T 0 transgenic plants were most readily obtained for both spring and winter wheat when mature embryo halves were used for co-inoculation by Agrobacterium cells. The presence of the antibiotic selection marker ( nptII , encoding neomycin phosphotransferase II) in the T 0 plants was revealed by both genomic PCR amplification and the enzyme-linked immunosorbent assay (ELISA). Additional analysis showed that the transgene was stably inherited from the two different generations and segregated normally among the T 1 progenies. Further development along this line will raise the efficiency of wheat transformation and increase the use of this approach in the molecular breeding of wheat crop.

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Aegiolops kotschyi cytoplasmic male sterile system often results in part of haploid plants in wheat (Triticum aestivum L.). To elucidate the origin of haploid, 235 wheat microsatellite (SSR) primers were randomly selected and screened for polymorphism between haploid (2n = 3x = 21 ABD) and its parents, male-sterile line YM21 (2n = 6x = 42 AABBDD) and male fertile restorer YM2 (2n = 6x = 42 AABBDD). About 200 SSR markers yielded clear bands from denatured PAGE, of which 180 markers have identifiable amplification patterns, and 20 markers (around 8%) resulted in different amplification products between the haploid and the restorer, YM2. There were no SSR markers that were found to be distinguishable between the haploid and the male sterile line YM21. In addition, different distribution of HMW-GS between endosperm and seedlings from the same seeds further confirmed that the haploid genomes were inherited from the maternal parent. After haploidization, 1.7% and 0.91% of total sites were up- and down-regulated exceeding twofold in the shoot and the root of haploid, respectively, and most of the differentially expressed loci were up/down-regulated about twofold. Out of the sensitive loci in haploid, 94 loci in the shoot, 72 loci in the root can be classified into three functional subdivisions: biological process, cellular component and molecular function, respectively.

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A recombinant inbred line (RIL) population with 302 lines derived from a cross of Weimai 8 × Luohan 2 was used to identify the quantitative trait loci (QTL) for plant height (PH) in wheat (Triticum aestivum L.). Possible genetic relationships between PH and PH components (PHC), including spike length (SL) and internode length from the first to the fourth node counted from the top, abbreviated as FIITL, SITL, TITL and FOITL, respectively, were evaluated at the QTL level. A QTL for PH was mapped using data on PH and on PH conditioned by PHC using the IciMapping V3.0 software. Conditional QTL mapping proved that, at the QTL level, SL contributed the least to PH, followed by FIITL and FOITL, while TITL had the strongest influence on PH, followed by SITL. These results indicate that the conditional QTL mapping method can be used to evaluate possible genetic relationships between PH and PHC, and that it can efficiently and precisely reveal counteracting QTL, which will enhance our understanding of the genetic basis of PH in wheat.

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Thinopyrum intermedium, which has many useful traits, is valuable for wheat breeding. A new wheat-Thinopyrum addition line, SN100109, was developed from the progeny of common wheat cultivar Yannong 15 and Th. intermedium. It was resistant to most races of Blumeria graminis f. sp tritici (Bgt), which caused powdery mildew in wheat, and its reactions were different from the reactions of gene Pm40 and Pm43. Genomic in situ hybridization (GISH) and molecular marker analysis were used to identify the genomic composition of SN100109. GISH results showed that SN100109 was a wheat-Th. intermedium disomic addition line containing one pair of J chromosomes, and the resistance gene was located on the alien additional chromosomes of SN100109. And four molecular markers BE425942, BF482714, Xgdm93 and BV679214 which were assigned to homologous group 2, were specific molecular markers of the additional chromosomes. All the results indicated that SN100109 contained one pair of 2J chromosomes. SN100109 can be used as a novel germplasm source for introducing powdery mildew resistance genes to wheat in breeding programs.

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High-yield common buckwheat ‘cv. Fengtian 1’ (FT1) and tartary buckwheat ‘cv. Jingqiao 2’ (JQ2) were selected to investigate the characteristics of the grain-filling process and starch accumulation of high-yield buckwheat. FT1 had an average yield that was 43.0% higher than that of the control ‘cv. Tongliaobendixiaoli’ (TLBDXL) in two growing seasons, while JQ2 had an average yield that was 27.3% higher than that of the control ‘cv. Chuanqiao 2’ (CQ2). The Richards equation was utilized to evaluate the grain-filling process of buckwheat. Both FT1 and JQ2 showed higher values of initial growth power and final grain weight and longer linear increase phase, compared with respective control. These values suggest that the higher initial increasing rate and the longer active growth period during grain filling play important roles to increase buckwheat yield. Similar patterns of starch, amylose and amylopectin accumulation were detected in common buckwheat, leading to similar concentration of each constituent at maturity in FT1 and TLBDXL. Tartary buckwheat showed an increasing accumulation pattern of amylose in developing seeds, which differed from that of starch and amylopectin. This pattern led to a significant difference of the concentrations of amylose and amylopectin at maturity between JQ2 and CQ2, the mechanisms of which remained unclear. Nevertheless, both FT1 and JQ2 showed increased starch, amylose, and amylopectin accumulation during the physiological maturity of grains. The results suggest that prolonging the active grain-filling period to increase carbohydrate partitioning from source to seed sink can be an effective strategy to improve buckwheat yield.

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Physiological male sterility induced by the chemical hybridizing agent (CHA) overcomes problems of maintenance of sterile lines and restorers. However, the mechanism of sterility is unclear. The process of tapetum of CHA-treated ‘Xi’nong 2611’ at uninucleate, binucleate and trinucleate were compared with control to determine if tapetum varying differently during developmental stages. Tapetal degradation in CHA-treated ‘Xi’nong 2611’ began at late uninucleate stage, somewhat earlier than control plants. Cytological observations indicated that the gradual degradation of the tapetum in CHA-treated ‘Xi’nong 2611’ was initiated and terminated earlier than in the control. These findings implied that CHA-induced male sterility was related to abnormally early tapetal degradation. In order to indicate the role of the SKP1 gene in fertility/sterility in wheat, its expression was assessed in anthers at uninucleate, binucleate and trinucleate stages. SKP1 expression was reduced in the later developmental stages, and there was an obvious decrease from the uninucleate to trinucleate stages. Higher expression of the SKP1 gene occurred in ‘Xi’nong 2611’ compared to CHA-treated ‘Xi’nong 2611’. This implied that SKP1 gene expression was inhibited during the fertility transformation process and was related to transformation from fertility to sterility. Moreover, the results from this study suggest that SKP1 plays an essential role of conducting fertility in physiological male sterility.

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Cereal Research Communications
Authors: N. Niu, Y.X. Bai, S. Liu, Q.D. Zhu, Y.L. Song, S.C. Ma, L.J. Ma, X.L. Wang, G.S. Zhang, and J.W. Wang

Studies of the pollen abortion mechanism in thermo-sensitive male sterile lines may provide a strong foundation for breeding hybrid wheat and establishing a theoretical basis for marker-assisted selection. To investigate the cause of pollen abortion in Bainong thermo – sensitive male sterile (BNS) lines, we analyzed the properties of pollen grains, changes in the tapetum and microspores in different anther developmental stages, and the distribution and deposition of nutrient substances in microspores. We found that tapetum degraded in the early uninucleate stage in sterile BNS (S-BNS), which was earlier than that of fertile BNS (F-BNS) tapetum. Large amounts of insoluble polysaccharides, lipids, and proteins were deposited until the trinucleate pollen stage in the nutritive cells in F-BNS. At the binucleate stage, the vacuoles disappeared and pollen inclusion increased gradually. At the trinucleate stage, these nutrients would help pollen grains mature and participate in fertilization normally. Therefore, early degradation of the tapetum, which inhibits normal microspore development, and the limited content of nutrient substances in pollen may be the main factors responsible for male sterility in BNS lines.

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Waxy wheat (Triticum aestivum L.) is grown throughout the world for its specific quality. Fertilization and planting density are two crucial factors that affect waxy wheat yield and photosynthetic capacity. The objectives of the research were to determine the effects of fertilization and planting density on photosynthetic characteristics, yield, and yield components of waxy wheat, including Yield, SSR, TGW, GNPP, GWPP, PH, HI, Pn, Gs, Ci, E and WUE using the method of field experiment, in which there were three levels (150, 300, and 450 kg ha−1) of fertilizer application rate and three levels (1.35, 1.8, and 2.25 × 106 plants ha−1) of planting density. The results suggested that photosynthetic characteristics, yield, and yield components had close relationship with fertilization levels and planting density. Under the same plant density, with the increase of fertilization, Yield, SSR, TGW, GNPP, GWPP, HI, Pn, Gs, E and WUE increased and then decreased, PH increased, but Ci decreased. Under the same fertilization, with the increase of plant density, Yield, SSR, TGW, GNPP, GWPP, HI increased and then decreased, PH, Pn, Gs and E increased, PH and WUE declined. The results also showed that F2 (300 kg ha−1) and D2 (1.8 × 106 plants ha−1) was a better match in this experiment, which could obtain a higher grain yield 4961.61 kg ha−1. Consequently, this combination of fertilizer application rate and plant densities are useful to get high yield of waxy wheat.

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Starch is a product of photosynthetic activities in leaves. Wheat yields largely depend on photosynthetic carbon fixation and carbohydrate metabolism in flag leaves. The mapping of quantitative trait loci (QTLs) associated with flag leaf starch content (FLSC) in wheat (Triticum aestivum L.) was completed using unconditional and conditional QTL analyses. The FLSC of this population during the early grain-filling stage was measured at six stages in six environments. Combining unconditional and conditional QTL mapping methods, eight unconditional QTLs and nine conditional QTLs were detected, with five QTLs identified as unconditional and conditional QTLs. Four unconditional QTLs (i.e. qFLS-1B, qFLS-1D-1, qFLS-4A, and qFLS-7D-1) and one conditional QTL (i.e. qFLS-3A-1) were identified in two of six environments. Two QTLs (qFLS-1D-2 and qFLS-7D-1), which significantly affected the FLSC, were identified using the unconditional QTL mapping method, while three QTLs (i.e. qFLS-1A, qFLS-3A-1, and qFLS-7D-1) were detected using the conditional QTL mapping method. Our findings provide new insights into the genetic mechanism and regulatory network underlying the diurnal FLSC in wheat.

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The aphid Sitobion avenae F. is one of the most harmful pests of wheat growth in the world. A primary field screening test was carried out to evaluate the S. avenae resistance of 527 wheat landraces from Shaanxi. The results indicated that 25 accessions (4.74%) were resistant to S. avenae in the three consecutive seasons, of which accession S849 was highly resistant, and seven accessions were moderately resistant. The majority of S. avenae resistant accessions come from Qinling Mountains. Then, the genetic variability of a set of 33 accessions (25 S. avenae resistant and 8 S. avenae susceptible) originating from Qinling Mountains have been assessed by 20 morphological traits and 99 simple sequence repeat markers (SSRs). Morphological traits and SSRs displayed a high level of genetic diversity within 33 accessions. The clustering of the accessions based on morphological traits and SSR markers showed significant discrepancy according to the geographical distribution, resistance to S. avenae and species of accessions. The highly and moderately resistant landrace accessions were collected from the middle and the east part of Qinling Mountains with similar morphology characters, for example slender leaves with wax, lower leaf area, and high ear density. These S. avenae resistant landraces can be used in wheat aphid resistance breeding as valuable resources.

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