Development of colchicine‐induced tetraploid St. Augustinegrass (Stenotaphrum secundatum) lines

St. Augustinegrass is well suited for lawns and commercial landscapes. While many genotypes are cross‐fertile, all cultivars are propagated vegetatively in sod production. To ensure varietal purity, development of sterile triploid hybrids by crossing tetraploid and diploid genotypes has been successfully used in other warm‐season turfgrasses. Applying this model in St. Augustinegrass would be beneficial to sod producers and turf managers who require purity for certification and uniformity for performance, respectively. This study was conducted to develop colchicine‐induced tetraploid lines of St. Augustinegrass. Seeds of cultivar ‘Raleigh’ were treated with four colchicine concentrations at four exposure times. A non‐treated control was included among the treatments. Seedlings that germinated were screened for genome size changes using flow cytometry. Line DSA 13005 and two progeny lines derived through selfing, DSA 16001 and DSA 16016, were corroborated as tetraploids (2n = 4x = 36) through chromosome counts. These lines will be used in future breeding efforts to attempt development of sterile triploid cultivars of St. Augustinegrass.     

寄主植物中抑制十字花科黑腐病菌Ⅲ型分泌系统小分子化合物的鉴定

 寻找抑制植物病原菌III型分泌系统的植物源活性小分子化合物,是研发生物安全农药的重要途径之一。本研究采用水煮提取法从十字花科黑腐病菌寄主植物满身红萝卜中提取分离活性小分子化合物,利用高效液相-质谱联用解析出活性物质的单体结构。然后用荧光素酶基因luxAB构建融合报告系统以及定量PCR检测活性物质对十字花科黑腐病菌III型分泌系统的抑制效果,最后采用剪叶接种和压渗接种的方法研究活性小分子物质对十字花科黑腐病菌的生防作用。研究表明,植物鞘氨醇和二氢鞘氨醇对十字花科黑腐病菌III型分泌系统基因的转录表达有一定程度的抑制作用,但是对I、II、IV型分泌系统基因的表达没有明显的抑制作用。植物鞘氨醇在XCM1上影响菌的生长,而二氢鞘氨醇不影响菌的生长。同时,还发现这两种物质能显著降低Xcc在寄主植株满身红萝卜上的病害症状以及能够使Xcc在非寄主植物辣椒上引起过敏反应的能力丧失。该研究结果为深入研究小分子化合物对十字花科黑腐病菌III型分泌系统的作用机制及后续开发植物源抑制剂提供了一定的理论依据。     

混合盐胁迫对马蔺幼苗根系构型及生长量的影响

为探讨不同种类盐胁迫对马蔺幼苗生长状况的影响,采用室内培养的方法,研究总质量浓度为0.3%的NaCl、NaHCO₃单盐溶液及其不同配比的混合盐溶液胁迫对马蔺(Iris lactea var. chinensis)幼苗生长量、地上部干鲜重比值、地下部干鲜重比值以及根系构型的影响。结果表明,随着胁迫时间的延长,0.3%NaCl溶液对叶数和株高增长量的影响最小,其次为3种混合盐溶液,各处理在第14天时株高增幅达到最大。经0.3%NaHCO₃处理后的幼苗地上部干鲜重的比值为空白对照的2.17倍,地下部干鲜重则为空白对照的56.61%,随着混合溶液中NaHCO₃浓度的上升,地上部干鲜重比值表现出上升的趋势,地下部干鲜重比值则表现出下降的趋势。经35天处理后,0.3%NaHCO₃胁迫的马蔺幼苗根系总长、根系表面积较对照均显著下降,降幅分别为39.73%、26.08%,而各处理间的根系体积却无显著差异。研究结果显示,0.3%NaHCO₃溶液对马蔺幼苗的胁迫作用最强,2种单盐对根系的胁迫作用强于混合盐胁迫,而0.3%NaCl溶液对生长量的胁迫作用最弱。     

Pyroptosis mediates high glucose-induced inflammation and injury in MC3T3-E1 osteoblasts

AIM To investigate whether pyroptosis contributes to the inflammation and injury in mouse embryonic osteoblastic cell line MC3T3-E1 induced by high glucose (HG; 45 mmol/L glucose). METHODS The cell viability was measured by CCK-8 assay. The protein expression levels of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) and caspase-1 (CASP1) were determined by Western blot. The secretion levels of interleukin-18 (IL-18) and IL-1β were measured by ELISA. The intracellular level of reactive oxygen species (ROS) was detected by 2',7'-dichlorodihydrofluorescein diacetate staining followed by photofluorography. Mitochondrial membrane potential (MMP) was examined by rhodamine 123 staining followed by photofluorography. The alkaline phosphatase (ALP) activity was determined using the ALP kit, and the number of mineralized nodules was detected by alizarin red S staining. RESULTS After the MC3T3-E1 osteoblasts were treated with HG for 24 h, the protein expression levels of NLRP3 and CASP1, and the secretion levels of IL-18 and IL-1β were significantly increased. The decrease in cell viability, and the increases in ROS generation and MMP loss were also observed. Moreover, the differentiation and mineralization of MC3T3-E1 osteoblasts were inhibited, evidenced by decreases in both ALP activity and mineralized nodule number. Knockdown of CASP1 by siRNA attenuated the HG-induced osteoblast inflammation and injury mentioned above. CONCLUSION Pyroptosis mediates HG-induced inflammation and injury in MC3T3-E1 osteoblasts.     

Mouse melanoma cell exosomes promote expression of Rac1 protein in fibroblasts

AIM To investigate the effect of exosomes secreted by mouse melanoma cells on the expression of Ras-related C3 botulinum toxin substrate 1 (Rac1) protein in fibroblasts. METHODS Ultracentrifugation was adopted to separete exosomes secreted by mouse melanoma B16-F10 cells. The morphological structure of exosomes was observed by negative-staining electron microscopy. The size distribution of exosomes was determined by nanoparticle tracking analysis (NTA). The exosomal markers, tumor susceptibility gene 101 (Tsg101) and tyrosinase-related protein 2 (Tyrp2), were identified by Western blot. Laser confocal microscopy was used to observe the process that mouse embryonic fibroblasts (MEF) took in exosomes during co-culture. Immunocytochemical staining and Western blot were used to detect the expression of Rac1 protein in MEF. RESULTS B16-F10 cell exosomes showed a typical tea tray-like structure, with a size range of 141~255 nm, and expressed protein markers Tsg101 and Tyrp2. The results of laser confocal microscopy showed that compared with co-culture at 0 h, a small number of exosomes appeared in the MEF at 12 h, and a large number of exosomes accumulated in the MEF after co-cultured for 24 and 36 h. Western blot analysis showed that compared with co-culture at 0 h, the expression of Rac1 protein in the MEF was significantly increased at 24 h and 36 h of co-culture (P<0.01). The results of immunocytochemical staining showed that compared with co-culture at 0 h, the positive expression level of Rac1 in the MEF cells was significantly increased at 12 h, 24 h and 36 h of co-culture (P<0.05 or P<0.01). CONCLUSION Intake of exosomes secreted by mouse melanoma cells promotes the expression of Rac1 protein in fibroblasts.     

Soluble Klotho protein suppresses THP-1-derived foam cell formation

AIMTo investigate the role of soluble Klotho protein in THP-1-derived foam cell formation. METHODSTHP-1 monocytes were induced into macrophages by treatment with 160 nmol/L phorbol myristate acetate for 48 h, and then were divided into 6 groups: negative control group (THP-1-derived macrophages), positive control group [THP-1-derived foam cells induced by oxidized low-density lipoprotein (ox-LDL) for 48 h], and 25, 50, 100 and 200 μg/L soluble Klotho protein groups (THP-1-derived macrophages pretreated with soluble Klotho protein at the indicat?ed concentraions for 2 h and then induced by ox-LDL for 48 h). Lipid droplets in cytoplasm were observed by oil red O staining. The cholesterol outflow rate was detected by scintillation counting technique. The content of intracellular total cholesterol, free cholesterol and cholesterol ester was detected by enzyme fluorescence analysis. The expression of acyl-coenzyme A:cholesterol acyltransferase 1 （ACAT1） and ATP-binding cassette transport?er A1 (ABCA1) at mRNA and protein levels was determined by RT-qPCR and Western blot, respectively. RESULTSOil red O staining and lipid mass quantification showed that THP-1-derived foam cell formation was dose-dependently suppressed by soluble Klotho protein. The cholesterol efflux rate of THP-1-derived foam cells was increased by soluble Klotho protein in a dose-dependent manner (P<0.05). In addition, soluble Klotho protein decreased the expression of ACAT1 and increased the expression of ABCA1 in a dose-dependent manner (P<0.05). CONCLUSION The soluble Klotho protein inhibits THP-1-derived foam cell formation in a dose-dependent manner by down-regulating the expression of ACAT1 and up-regulating the expression of ABCA1.     

Role of IP3R in LH-EGFR-induced mouse oocyte meiotic resumption

AIM: To explore the effect of inositol 1, 4, 5-trisposphate receptor (IP₃R) in luteinizing hormone-epidermal growth factor receptor (LH-EGFR)-induced oocyte meiotic resumption. METHODS: Models of mouse cumulus-oocyte complexs (COCs) culture and follicle culture in vitro were generated to study the effects of 2-aminoethyl diphenyl borate (2-APB) and heparin (IP₃R specific inhibitors) on LH/EGF-induced oocyte meiotic resumption and EGF-induced cumulus cell expansion. Real-time PCR was used to detect the mRNA expression of cumulus expansion-related factors. The changes of the intracellular calcium level were monitored using Fluo 3-AM, and the cGMP level was measured by ELISA. RESULTS: The inhibitors of IP₃R, 2-APB and heparin, dramatically reversed EGF-induced oocyte maturation (P<0.05) and decreased cGMP levels in COCs (P<0.05). In addition, 2-APB and heparin reversed EGF-induced cumulus expansion, and significantly inhibited EGF-induced cumulus expansion-related factor expression (P<0.05). The activation of IP₃R increased intracellular calcium level, and the study found that 2-APB and heparin dramatically reversed EGF-induced elevation of calcium level in cumulus cells (P<0.05). Follicular culture in vitro showed that 2-APB and heparin significantly reversed the LH-induced oocyte maturation (P<0.05). CONCLUSION: LH-EGFR signaling pathway increases calcium level in cumulus cells through IP₃R, resulting in meiotic resumption.     

四川地区猕猴桃病毒1的RT-PCR检测及外壳蛋白基因序列分析

为了明确近期新西兰报道的侵染猕猴桃的新病毒——猕猴桃病毒1（Actinidia virus 1，AcV-1）在四川地区猕猴桃上的发生情况及其分子特性，采用RT-PCR对来自四川6个地区疑似感染病毒的90份猕猴桃主栽品种‘红阳’和‘金果’的叶片样品进行检测。结果表明，22份样品为AcV-1阳性，检出率为24.4%。将获得的5个AcV-1分离物和已报道的新西兰分离物K75的外壳蛋白（coat protein，CP）基因序列进行比对，结果表明，分离物间核苷酸序列和氨基酸序列的相似性分别为84.8% ~ 97.1%和89.7% ~ 99.6%，其中除邛崃分离物HYH5与新西兰分离物K75的核苷酸序列相似性较高（97.1%）外，其余4个分离物均低于91%。系统进化分析结果显示，这些分离物主要聚集在3个分支上，分离物HYH5与新西兰分离物K75位于同一分支，其余分离物位于另外2个分支。     

羊角椒新品种寿研梦扬的选育

寿研梦扬是由母本CSVHPM1407001和父本CSVHPF1407002配制而成的羊角椒一代杂种。植株长势旺盛,连续坐果能力强;果实长羊角形,果长25～30 cm,果肩宽3～4 cm;青熟果黄绿色,老熟果红色;外表光亮,商品性好,耐贮运;单果质量100 g左右,辣味浓,宜鲜食;田间对病毒病、炭疽病和疫病的抗性强于对照喜洋洋。保护地栽培鲜椒产量可达12 840 kg·(667 m~2)~(-1)左右。适宜山东、河北等地区早春、秋延保护地种植。     

黑甜糯玉米新品种黑甜糯639的选育

黑甜糯639是以糯玉米自交系HNF为母本,以甜玉米自交系D91为父本配制而成的黑甜糯玉米一代杂种。生育期为91 d(天),株高263 cm,穗位151 cm,雄穗主轴与分枝角度中等,一级分枝18～23个,果穗筒型,籽粒黑紫色,甜糯籽粒比例为1∶3,籽粒可溶性糖含量为12.5%,支链淀粉占总淀粉含量的98.1%,口感佳。高抗丝黑穗病、茎腐病、大斑病,抗穗腐病和矮花叶病,平均每667 m~2鲜穗产量900 kg左右。适宜在山西省鲜食糯玉米主产区种植。