60Co-γ射线和电子束辐照灭菌对葛根提取物抗氧化活性及指纹图谱的影响

为评价⁶⁰Co-γ射线和电子束辐照灭菌对葛根提取物品质的影响效应及其差异性,采用不同剂量的⁶⁰Co-γ射线(0、2.8、5.7、8.3、10.6 kGy)和电子束(0、2.4、5.1、7.7、10.3 kGy)辐照处理葛根提取物,研究辐照对其微生物总数、抗氧化活性及指纹图谱的影响。结果表明,⁶⁰Co-γ射线和电子束辐照均能有效降低葛根提取物中需氧菌总数、霉菌和酵母菌总数,随着吸收剂量增大,抑制作用增强。⁶⁰Co-γ射线和电子束辐照吸收剂量分别为5.7 kGy和5.1 kGy时,葛根提取物均达到《中国药典》所规定的微生物限度标准(需氧菌总数为10³ CFU·g^-1、霉菌和酵母菌总数为10² CFU·g^-1);吸收剂量分别为5.7 kGy和7.7 kGy时,需氧菌总数、霉菌和酵母菌总数均降至10 CFU·g^-1以下。此外,大肠埃希氏菌(Escherichia coli)与沙门氏菌(Salmonella)均未检出。2种辐照方式均对葛根提取物的DPPH自由基(DPPH·)清除率、羟自由基(·OH)清除率以及总还原能力无显著影响(P>0.05)。不同剂量⁶⁰Co-γ射线和电子束辐照处理葛根提取物特征峰的相似度达到1.0,且相对保留时间的相对标准差(RSD)均小于0.5%,表明2种辐照方式对葛根提取物指纹图谱均无显著影响(P>0.05)。综上所述,⁶⁰Co-γ射线和电子束辐照对葛根提取物微生物具有较强的抑制作用,2种辐照方式均不会对葛根提取物的抗氧化活性及指纹图谱产生显著影响,不影响葛根提取物的有效性及质量稳定性。该研究为2种辐照技术,特别是电子束辐照在葛根药材、饮片及其中成药制剂灭菌中的合理应用提供了科学参考。     

Electron Transport Through Photosystem II Is Not Limited By A Wide Range of Water Deficit Conditions In Field‐Grown Gossypium hirsutum

Despite exhaustive literature describing drought stress effects on photosynthesis in Gossypium hirsutum, the sensitivity of photosynthetic electron flow to water deficit is heavily debated. To address this, G. hirsutum plants were grown at a field site near Camilla, GA under contrasting irrigation regimes, and pre‐dawn water potential (Ψ_PD), stomatal conductance (g_s), net photosynthesis (P_N), actual quantum yield of photosystem II (Φ_PSII) and electron transport rate (ETR) were measured at multiple times during the 2012 growing season. Ψ_PD values ranged from ?0.3 to ?1.1 MPa. Stomatal conductance exhibited a strong (r² = 0.697), sigmoidal response to Ψ_PD, where g_s was ≤0.1 mol m^?2 s^?1 at Ψ_PD values ≤ ?0.86 MPa. Neither Φ_PSII (r² = 0.015) nor ETR (r² = 0.010) was affected by Ψ_PD, despite exceptionally low Ψ_PD values (?1.1 MPa) causing a 71.7 % decline in P_N relative to values predicted for well‐watered G. hirsutum leaves at Ψ_PD = ?0.3 MPa. Further, P_N was strongly influenced by g_s, whereas ETR and Φ_PSII were not. We conclude that photosynthetic electron flow through photosystem II is insensitive to water deficit in field‐grown G. hirsutum.     

小反刍兽疫病毒及山羊痘病毒N-ITR基因二联实时荧光定量RT-PCR检测方法的建立

Peste des petits ruminants virus (PPRV) and goat pox virus (GTPV) are the causative agents of two kinds of goats’ diseases-peste des petits ruminants and goat pox which can cause disaster economic losses. In order to detect the two viruses simultaneously and quickly, two sets of primers and relative probes were designed based on the nucleoprotein (N) gene of PPRV and the inverted terminal repeat (ITR) segment of GTPV， respectively. In order to work together in the same reaction, the probes were labeled with different fluorescent materials 5′FAM-TAMRA3′and 5′JOE-Eclipse3′，respectively. Results showed that the duplex Real-time RT-PCR assay was identified to be specific for PPRV and GTPV only and specific fluorescent signal could be detected, but the related viruses including fowl pox virus（FPV）and canine distemper virus (CDV) had no specific fluorescent signal. Positive recombinant plasmids (PPRV pMD18-T-N and GTPV pMD18-T-ITR) were built and used for positive quantitative templates to establish duplex standard curves. The developed assay based on the probe N-ITR was found to be highly specific and sensitive with a detection limit of 10² copies/μL cDNA and 10³ copies/μL DNA for PPRV and GTPV， respectively. Finally, the duplex Real-time RT-PCR assay for simultaneous detection of PPRV and GTPV was established preliminarily in the study.     

强闪光抑制棉花叶片光系统Ⅱ活性和热耗散

除持续强光导致光合作用效率降低外,强闪光也能够影响光合功能,但规律和机制尚不清楚。为研究强闪光对喜光植物棉花叶片光合功能的影响,选用陆地棉(Gossypium hirsutum L.)品种新陆早45号为材料,于强闪光处理(20,000μmol m–2 s–1, 300 ms,间隔10 s,处理时间持续30 min)前后分别测定叶绿素荧光、P700和气体交换。结果表明,强闪光处理后不仅有活性的PSI (光系统I)反应中心含量下降,同时PSⅡ (光系统Ⅱ)电子传递活性也受到限制。与对照相比,强闪光处理后PSI的ΦND (PSI供体侧限制引起的非光化学量子产量)下降,ΦNA (PSI受体侧限制引起的非光化学量子产量)增加,暗示强闪光能够抑制PSI受体侧电子传递活性。强闪光处理不仅使PSⅡ的实际量子产量明显下降,而且非光化学猝灭和ΦNPQ (PSⅡ调节性能量耗散的量子产量)也降低;但是,ΦNO (PSⅡ非调节性能量耗散的量子产量)明显增加,表明强闪光导致热耗散降低和PSⅡ失活。此外,强闪光处理后光合速率和气孔导度均降低,但细胞间隙CO2浓度增加,证明强闪光处理后同化能力的降低不是气孔限制导致的。因此,本研究认为强闪光处理不仅抑制PSI活性,而且导致PSⅡ失活和可调节性热耗散下降;光合电子传递活性的下降可能是强闪光下光合速率降低的重要原因。     

Development of Nucleic Acid Lateral Flow Immunoassay Detection Method for Foot-and-mouth Disease Virus

In this article, through the combination of nucleic acid probes and immune chromatography, a simple, sensitive and specific detection system——nucleic acid lateral flow immunoassay (NALFIA) for amplifing foot-and-mouth disease virus (FMDV) 3D RT-PCR products was established.An ultrasensitive nucleic acid biosensor (NAB) based on streptavidin-labeled gold nanoparticles dual labels and lateral flow strip biosensor (LFSB) were used in this system.The biotinylated goat anti-rabbit IgG was marked to the NC membrane as the alleged strip and the anti-digoxin antibody was labeled to the NC membrane to capture the digoxin probe.After assemblying gold-labeled strip and detecting RT-PCR products, the detection limit of NALFIA was 0.3×10^-3 to 3×10^-3 μg/μL.The NALFIA was compared with agar gel electrophoresis analysis, the results showed that the sensitivity of NALFIA was higher than agar gel electrophoresis.There was an excellent agreement between the two methods.NALFIA was a method with high sensitive, low cost and short time.In conclusion, this method provided a good alternative to detect FMDV.     

More evidence that anaerobic oxidation of methane is prevalent in soils: Is it time to upgrade our biogeochemical models?

Estimating future fluxes of CH₄ between land and atmosphere requires well-conceived process-based biogeochemical models. Current models do not represent the anaerobic oxidation of methane (AOM) in land surface soils, in spite of increasing evidence that this process is widespread. Our objective was to determine whether AOM, or potential AOM, commonly occurs in 20 hydromorphic soils spanning a wide range of chemical properties. Bulk soil samples were collected under shallow water near the shoreline of 15 recently drained fish ponds in southern Bohemia (Czech Republic), as well as from below the water table at 3 peatland locations in northeast Scotland and 2 acid sulfate soils on the southern coast of Finland. Each soil slurry was incubated under both oxic and anoxic conditions, with or without the addition of alternative electron acceptors (SO₄²⁻ and NO₃⁻) or H₂PO₄⁻. Here, “oxic” and “anoxic” conditions refer to anoxic soil respectively incubated in a headspace containing air or argon. Using the isotope dilution method, we determined the gross production and oxidation rates of CH₄ after 2 days incubation under oxic headspace conditions, and after 2, 21 and 60 days incubation under anoxic conditions. Large differences in net CH₄ fluxes were observed between soil types and between incubation conditions. AOM was detected in each of the 20 bulk soil samples, which spanned >6 pH units and 2 orders of magnitude in organic C content. Significant positive relationships were found between AOM and gross CH₄ production rates under anoxic conditions, resulting in AOM rates that were sometimes higher than CH₄ oxidation rates under oxic headspace conditions. There was no relationship between net and gross CH₄ production rates, such that 2 soil types could display similar low net rates, yet conceal very large differences in gross rates. The effects of alternative electron acceptors on AOM were idiosyncratic and resulted in no net trend. We did find, however, a negative effect of SO₄²⁻ and H₂PO₄⁻ on gross CH₄ production rates under anoxic and oxic conditions respectively. Under oxic headspace conditions, CH₄ oxidation was related to soil organic C content. Taken collectively, our results suggest that AOM, or potential AOM, is prevalent over a wide range of soil types, that AOM may contribute substantially to CH₄ oxidation in soils, and that AOM in soils should be integrated to current process-based CH₄ cycling models.     

Histological and proteomics analysis of apple defense responses to the development of Colletotrichum gloeosporioides on leaves

Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. is an important fungal pathogen known to cause glomerella leaf spot (GLS). The objective of this study was to analyze the infection of C. gloeosporioides on leaves of apple cv. Fuji (resistant) and cv. Gala (susceptible) and apply proteomics techniques to study the apple defense responses 48 h after inoculation (h.a.i.). On both of cultivars, C. gloeosporioides started to germinate at 3 h.a.i. on adaxial surface and produced appressoria adhering to epidermal cell juxtapositions. Histological analysis showed more stratified parenchyma in leaves of cv. Fuji than cv. Gala associated with differences in the chemical composition of cell walls. Total and unique proteins expressed by cvs. Fuji and Gala at 3, 12, 24 and 48 h.a.i. were detected by comparative proteomes analysis. A total of 42 unique proteins expressed at 24 and 48 h.a.i. were identified by MALDI/TOF mass spectrometry, and most of these proteins were identified as directly involved in defense responses.     

棉花花铃期低温对叶片PSI和PSII光抑制的影响

选用陆地棉(Gossypium hirsutum L.)品种新陆早45号,在室外盆栽至开花结铃期后,移至人工气候室,模拟新疆棉花花铃期易出现的低温逆境条件,设置处理T(16℃/10℃,昼/夜),以常温(30℃/18℃,昼/夜)处理作为对照,采用叶绿素荧光和P700同步测定技术,研究低温对棉花花铃期叶片光合机构PSII能量分配、PSI氧化还原状态及环式电子传递流的影响。结果表明,与对照相比,低温处理显著降低了棉花叶片PSII光适应状态下最大光化学量子产量(F_v'/F_m')、光化学猝灭系数(qP)和PSII有效光化学量子产量[Y(II)],并使PSII非调节性能量耗散[Y(NO)]和调节性能量耗散[Y(NPQ)]量子产量显著升高,诱导PSII发生光抑制。低温引起棉花叶片光合机构PSI受体侧限制[Y(NA)]显著下降和供体侧限制[Y(ND)]显著升高,但未引起有效的PSI复合体含量(P_m)显著降低,表明与PSII相比,棉花叶片PSI对低温不敏感。此外,低温引起环式电子传递量子产量[Y(CEF)]以及与PSII实际量子产量比率的[Y(CEF)/Y(II)]显著升高,进一步表明在低温下,光破坏防御机制中环式电子传递流对棉花PSI、PSII起着重要的保护作用,是主要的光破坏防御机制。非光化学热耗散(NPQ)和调节性非光化学热耗散[Y(NPQ)]与[Y(CEF)]具有显著的正相关关系,表明低温引起棉花花铃期叶片PSII反应中心过度关闭产生过剩的激发能,造成了PSII可逆的光抑制,环式电子传递流的响应及较高的调节性能量耗散共同保护了棉花叶片PSI和PSII免受光抑制的损伤,这可能是棉花叶片PSI对低温不敏感的重要原因。     

不同固定条件对几种植物样品超微结构的影响

为比较几种固定条件对不同植物材料样品电镜超微结构图像清晰度的影响。以3个不同科属4种植物[油棕(Elaeis gineansis Jacq.)、拟南芥(Arabidopsis thaliana.)、花生(Arachis hypogaea Linn.)、柱花草(Stylosanthes guianensias Sw.)]为研究主体代表,探讨了2.5%戊二醛与4%戊二醛固定过夜、锇酸固定2 h及过夜的制备方法对其叶片超微结构的影响,并成功观察了柱花草接种胶苞炭疽野生型菌株CH008后的病菌侵入过程。结果表明:油棕、拟南芥、花生及柱花草叶片经4%戊二醛与1%锇酸固定液固定过夜,其组织成像的效果均较为理想,各种细胞器整体信息丰富、结构组织完好、线条清晰。表明,此制备方法能够较好的保存植物叶片和细胞组织,提高制样的成功率及观察分辨率。     

红素氧还蛋白研究进展

自然界中大多数的红素氧还蛋白(rubredoxin)集中来自严格厌氧菌,包括细菌、古菌和少数微需氧细菌,而植物中该类蛋白相对较少,它的功能主要表现在作为辅因子或电子供体参与多个生化途径。也有研究显示,无论是植物中还是微生物中的红素氧还蛋白基因在协助生物抵御氧化胁迫方面均发挥着重要的作用。本文全面介绍自然界中的红素氧还蛋白基因的分布、结构以及在微生物、植物中所发挥的作用等,希望能为今后的进一步研究提供参考。