Isolation and characterization of a wheat--Psathyrostachys huashanica 'Keng' 3Ns disomic addition line with resistance to stripe rust.

We isolated a wheat germplasm line, 22-2, which was derived from common wheat (Triticum aestivum '7182') and Psathyrostachys huashanica 'Keng' (2n = 2x = 14, NsNs). Genomic composition and homoeologous relationships of 22-2 was analyzed using cytology, genomic in situ hybridization (GISH), EST-SSR, and EST-STS to characterize the alien chromatin in the transfer line. The cytological investigations showed that the chromosome number and configuration were 2n = 44 = 22 II. Mitotic and meiotic GISH using P. huashanica genomic DNA as the probe indicated that 22-2 contained a pair of P. huashanica chromosomes. The genomic affinities of the introduced P. huashanica chromosomes were determined by EST-SSR and EST-STS using multiple-loci markers from seven wheat homoeologous groups between the parents and addition line. One EST-SSR and 17 EST-STS markers, which were located on the homoeologous group 3 chromosomes of wheat, amplified polymorphic bands in 22-2 that were unique to P. huashanica. Thus, these markers suggested that the introduced Ns chromosome pair belonged to homoeologous group 3, so we designated 22-2 as a 3Ns disomic addition line. Based on disease reaction to mixed races (CYR31, CYR32, and Shuiyuan14) of stripe rust in the adult stages, 22-2 was found to have high resistance to stripe rust, which was possibly derived from its P. huashanica parent. Consequently, the new disomic addition line 22-2 could be a valuable donor source for wheat improvement depending on the excellent agronomic traits, especially, the introduction of novel disease resistance genes into wheat during breeding programs.

QTL mapping and genomic prediction of resistance to wheat head blight caused by Fusarium verticillioides.

Fusarium head blight (FHB), is one of the destructive fugue diseases of wheat worldwide caused by the Fusarium verticillioides (F.v). In this study, a population consisting of 262 recombinant inbred lines (RILs) derived from Zhongmai 578 and Jimai 22 was used to map Quantitative Trait Locus (QTL) for FHB resistance, with the genotype data using the wheat 50 K single nucleotide polymorphism (SNP) array. The percentage of symptomatic spikelet (PSS) and the weighted average of PSS (PSSW) were collected for each RIL to represent their resistance to wheat head blight caused by F.v. In total, 22 QTL associated with FHB resistance were identified on chromosomes 1D, 2B, 3B, 4A, 5D, 7A, 7B, and 7D, respectively, from which 10 and 12 QTL were detected from PSS and PSSW respectively, explaining 3.82%-10.57% of the phenotypic variances using the inclusive composite interval mapping method. One novel QTL, Qfhb. haust-4A.1, was identified, explaining 10.56% of the phenotypic variation. One stable QTL, Qfhb. haust-1D.1 was detected on chromosome 1D across multiple environments explaining 4.39%-5.70% of the phenotypic variation. Forty-seven candidate genes related to disease resistance were found in the interval of Qfhb. haust-1D.1 and Qfhb. haust-4A.1. Genomic prediction accuracies were estimated from the five-fold cross-validation scheme ranging from 0.34 to 0.40 for PSS, and from 0.34 to 0.39 for PSSW in in-vivo inoculation treatment. This study provided new insight into the genetic analysis of resistance to wheat head blight caused by F.v, and genomic selection (GS) as a potential approach for improving the resistance of wheat head blight.

Clinical observation on the treatment of childhood refractory idiopathic thrombocytopenic purpura with Dihuang Zhixue Capsule.

OBJECTIVE: To observe the clinical effect of Dihuang Zhixue Capsule (DZC, a Chinese preparation for cooling blood and dispelling toxic substances) in the treatment of childhood refractory idiopathic thrombocytopenic purpura (RITP), with cyclosporin A (CsA) used as the control. METHODS: Forty-one children of RITP were randomized into the treated group and the control group. The 21 patients in the treated group were orally given 2 to 3 DZC capsules each time, thrice a day and the 20 in the control group were given 3 mg/kg CsA per day, with 3 months as one therapeutic course. The therapeutic efficacy, platelet count and adverse reaction in the two groups were compared at the end of the course. RESULTS: (1) In the treated group, 1 (4.8%) patient was evaluated as cured, 3 (14.3%) as markedly effective, 5 (23.8%) as effective, 5 (23.8%) as improved, 7 (33.3%) as ineffective, with the total effective rate being 66.7%; while in the control group, the corresponding numbers were 0, 2 (10.0%), 2 (10.0%), 3 (15.0%), 13 (65.0%) and 35.0%, respectively, showing statistical significance in difference between the total effective rates of the two groups (xi(2)=4.11, P=0.0426). (2) As compared with the baseline, the platelet count increased in both groups after 2 months' treatment (P<0.05). After 3 months' treatment, the platelet count was higher in the treated group than in the control group (P<0.05). (3) The improvement of hemorrhage in the treated group after 8 weeks' treatment was better than that in the control group (P<0.05). (4) No apparent adverse reaction was observed in the treated group, while in the control group, hirsutism was shown in 15 cases; gingival hyperplasia in 10; digestive reaction in 5, liver function impairment in 5, hypertension in 2 and renal impairment in 2. CONCLUSION: The therapeutic efficacy of DZC is better than that of CsA, and DZC shows good compliance but brings no obvious adverse reaction.

ClC5 Decreases the Sensitivity of Multiple Myeloma Cells to Bortezomib via Promoting Prosurvival Autophagy.

Resistance to bortezomib (BZ) is the major problem that largely limits its clinical application in multiple myeloma treatment. In the current study, we investigated whether ClC5, a member of the chloride channel family, is involved in this process. The MTT assay showed that BZ treatment decreased cell viability in three multiple myeloma cell lines (ARH77, U266, and SKO-007), with IC50 values of 2.83, 4.37, and 1.91 nM, respectively. Moreover, BZ increased the conversion of LC3B-I to LC3B-II and expressions of beclin-1 and ATG5, concomitantly with a decreased p62 expression. Pharmacological inhibition of autophagy with 3-MA facilitated cell death in response to BZ treatment. Additionally, BZ increased ClC5 protein expression in ARH77, U266, and SKO-007 cells. Knockdown of ClC5 with small interfering RNA sensitized cells to BZ treatment, and upregulation of ClC5 induced chemoresistance to BZ. Furthermore, ClC5 downregulation promoted BZ-induced LC3B-I to LC3B-II conversion and beclin-1 expression, whereas overexpression of ClC5 showed the opposite results in ARH77 cells. Finally, BZ induced dephosphorylation of AKT and mTOR, which was significantly attenuated by ClC5 inhibition. However, ClC5 upregulation further enhanced AKT and mTOR dephosphorylation induced by BZ. Our study demonstrates that ClC5 induces chemoresistance of multiple myeloma cells to BZ via increasing prosurvival autophagy by inhibiting the AKT-mTOR pathway. These data suggest that ClC5 may play a critical role in future multiple myeloma treatment strategies.

Molecular cytogenetic characterization of a wheat - Leymus mollis 3D(3Ns) substitution line with resistance to leaf rust.

Leymus mollis (Trin.) Pilger (NsNsXmXm, 2n = 28), a wild relative of common wheat, possesses many potentially valuable traits that could be transferred to common wheat during breeding programs. In this study, the karyotypic constitution of a wheat - L. mollis 3D(3Ns#1) disomic substitution line isolated from the F5 progeny of octoploid Tritileymus M842-16 × Triticum durum cv. D4286, which was designated as 10DM57, was determined using genomic in situ hybridization (GISH), fluorescent in situ hybridization (FISH), SSR markers, and EST-STS markers. Screening of mitosis and meiosis showed that 10DM57 had a chromosome karyotype of 2n = 42 = 21II. GISH indicated that 10DM57 was a line with 40 chromosomes from wheat and two of the Ns chromosomes from L. mollis, which formed a ring bivalent in pollen mother cells at metaphase I. FISH analysis showed that the chromosome 3D may be replaced by 3Ns#1 in 10DM57. DNA markers, including SSR and EST-STS primers, showed that the pair of wheat chromosome 3D in 10DM57 was substituted by the pair of chromosome 3Ns#1 from L. mollis. Evaluation of the agronomic traits showed that, compared with its common wheat relative 7182, 10DM57 was resistant to leaf rust while the spike length and number of spikes per plant were improved significantly, which correlated with a higher wheat yield. The new germplasm, 10DM57, could be exploited as an intermediate material in wheat genetic and breeding programs.

Isolation and characterization of a Psathyrostachys huashanica Keng 6Ns chromosome addition in common wheat.

The development of alien addition lines is important for transferring useful genes from exotic species into common wheat. A hybrid of common wheat cv. 7182 (2n = 6x = 42, AABBDD) and Psathyrostachys huashanica Keng (2n = 2x = 14, NsNs) via embryo culture produced the novel intergeneric disomic addition line 59-11. The seed morphology of 59-11 resembled the parent 7182 and it exhibited extreme agronomic characteristics, i.e., twin stable spikelets, fertile florets, and multi-kernel clusters. Furthermore, 59-11 produced plump kernels with a high seed-setting percentage during the advanced maturation stage. The line was screened based on genomic in situ hybridization, EST-SSR, EST-STS, and gliadin to identify P. huashanica chromatin in the wheat background. The chromosome number and configuration of 59-11 was 2n = 44 = 22 II and we confirmed the 6Ns disomic chromosome additions based on A-PAGE analysis and molecular markers. The results suggested that the production of twin spikelets and multiple kernels per spike in the wheat-P. huashanica addition line was related to homologous group 6 in the wheat chromosome. This is the first report of the introduction of improved spike traits into common wheat from the alien species P. huashanica and it opens up the possibility of increasing the wheat yield based on this enlarged gene pool.