Recording of calcium transient and analysis of calcium removal mechanisms in cardiac myocytes from rats and ground squirrels

With confocal microscopy, we recorded calcium transients and analyzed calcium removal rate at different temperatures in cardiac myocytes from the rat, a non-hibernator, and the ground squirrel, a hibernator. The results showed a remarkable increase of the diastolic level of calcium transients in the rat but no detectable change in the ground squirrel. Calcium transient of the ground squirrel, compared with that of the rat at the same temperature, had a shorter duration and showed a faster calcium removal. As indicated by the pharmacological effect of cyclopiazonic acid, calcium uptake by sarcoplasmic reticulum (SR) was the major mechanism of calcium removal, and was faster in the ground squirrel than in the rat. Our results confirmed the essential role of SR in hypothermia-tolerant adaptation, and negated the importance of Na-Ca exchange. We postulated the possibility to improve hypothermia-tolerance of the cardiac tissue of non-hibernating mammals.     

同域分布的珍稀野生动物对放牧的行为响应策略

保护区内放牧活动对野生动物保护存在负面影响,明确不同物种对放牧干扰的行为响应对制定更有针对性的保护管理政策具有重要意义。使用红外相机研究卧龙自然保护区放牧活动对多种珍稀野生动物的影响,分析放牧激励政策实施前后大熊猫(Ailuropoda melanoleuca)及其同域分布的小熊猫(Ailurus fulgens)、川金丝猴(Rhinopithecus roxellana)、水鹿(Rusa unicolor) 4种珍稀野生动物的照片数、空间分布以及活动模式的变化,探讨这4种动物对放牧的行为响应策略。结果表明:(1)一期(2012—2013),2012年10月实施了禁马政策,同年12月实施放牧(牛羊)激励政策)家畜照片数量很少,4种野生动物照片数相对较多。二期(2014—2015)家畜的照片数显著增加(P0.01),小熊猫(P0.05)与川金丝猴(P0.01)的照片数均显著减少,大熊猫、水鹿的照片数也呈减少趋势;到三期(2016—2017),大熊猫、小熊猫及水鹿3种关注野生动物的照片数基本回升到激励政策实施前的水平,无川金丝猴照片记录。(2)一期,4种野生动物在研究区域有较广的分布;二期,大熊猫、小熊猫的空间分布范围均缩小,无川金丝猴空间分布信息,而家畜、水鹿的空间分布范围有所增加;到三期,大熊猫、小熊猫的空间分布基本恢复到放牧激励政策实施前的区域,无川金丝猴的空间分布信息。(3)放牧激励政策实施前后,大熊猫、小熊猫及川金丝猴活动模式无明显变化,但水鹿的活动更加集中于傍晚,以避开人类与家畜的活动高峰。同域分布的不同的野生动物对人类活动(如放牧)的行为响应策略不同,各保护区在制定相关保护政策时应综合考虑人类干扰对多个物种的影响,增加决策的科学性与合理性。     

Plant nitrogen dynamics and nitrogen-use strategies under altered nitrogen seasonality and competition

BACKGROUND AND AIMS: Numerous studies have examined the effects of climatic factors on the distribution of C(3) and C(4) grasses in various regions throughout the world, but the role of seasonal fluctuations in temperature, precipitation and soil N availability in regulating growth and competition of these two functional types is still not well understood. This report is about the effects of seasonality of soil N availability and competition on plant N dynamics and N-use strategies of one C(3) (Leymus chinensis) and one C(4) (Chloris virgata) grass species. METHODS: Leymus chinensis and C. virgata, two grass species native to the temperate steppe in northern China, were planted in a monoculture and a mixture under three different N seasonal availabilities: an average model (AM) with N evenly distributed over the growing season; a one-peak model (OM) with more N in summer than in spring and autumn; and a two-peak model (TM) with more N in spring and autumn than in summer. KEY RESULTS: The results showed that the altered N seasonality changed plant N concentration, with the highest value of L. chinensis under the OM treatment and C. virgata under the TM treatment, respectively. N seasonality also affected plant N content, N productivity and N-resorption efficiency and proficiency in both the C(3) and C(4) species. Interspecific competition influenced N-use and resorption efficiency in both the C(3) and C(4) species, with higher N-use and resorption efficiency in the mixture than in monoculture. The C(4) grass had higher N-use efficiency than the C(3) grass due to its higher N productivity, irrespective of the N treatment or competition. CONCLUSIONS: The observations suggest that N-use strategies in the C(3) and C(4) species used in the study were closely related to seasonal dynamics of N supply and competition. N seasonality might be involved in the growth and temporal niche separation between C(3) and C(4) species observed in the natural ecosystems.     

Is Mitochondrial tRNAphe Variant m.593T>C a Synergistically Pathogenic Mutation in Chinese LHON Families with m.11778G>A?

Mitochondrial transfer RNA (mt-tRNA) mutations have been reported to be associated with a variety of diseases. In a previous paper that studied the mtDNA background effect on clinical expression of Leber''s hereditary optic neuropathy (LHON) in 182 Chinese families with m.11778G>A, we found a strikingly high frequency (7/182) of m.593T>C in the mitochondrially encoded tRNA phenylalanine (MT-TF) gene in unrelated LHON patients. To determine the potential role of m.593T>C in LHON, we compared the frequency of this variant in 479 LHON patients with m.11778G>A, 843 patients with clinical features of LHON but without the three known primary mutations, and 2374 Han Chinese from the general populations. The frequency of m.593T>C was higher in LHON patients (14/479) than in suspected LHON subjects (12/843) or in general controls (49/2374), but the difference was not statistically significant. The overall penetrance of LHON in families with both m.11778G>A and m.593T>C (44.6%) was also substantially higher than that of families with only m.11778G>A (32.9%) (P = 0.083). Secondary structure prediction of the MT-TF gene with the wild type or m.593T>C showed that this nucleotide change decreases the free energy. Electrophoretic mobility of the MT-TF genes with the wild type or m.593T>C transcribed in vitro further confirmed the change of secondary structure in the presence of this variant. Although our results could suggest a modest synergistic effect of variant m.593T>C on the LHON causing mutation m.11778G>A, the lack of statistical significance probably due to the relatively small sample size analyzed, makes necessary more studies to confirm this effect.     

Optogenetic manipulation of cyclic guanosine monophosphate to probe phosphodiesterase activities in megakaryocytes

Cyclic guanosine monophosphate (cGMP) signalling plays a fundamental role in many cell types, including platelets. cGMP has been implicated in platelet formation, but mechanistic detail about its spatio-temporal regulation in megakaryocytes (MKs) is lacking. Optogenetics is a technique which allows spatio-temporal manipulation of molecular events in living cells or organisms. We took advantage of this method and expressed a photo-activated guanylyl cyclase, Blastocladiella emersonii Cyclase opsin (BeCyclop), after viral-mediated gene transfer in bone marrow (BM)-derived MKs to precisely light-modulate cGMP levels. BeCyclop-MKs showed a significantly increased cGMP concentration after illumination, which was strongly dependent on phosphodiesterase (PDE) 5 activity. This finding was corroborated by real-time imaging of cGMP signals which revealed that pharmacological PDE5 inhibition also potentiated nitric oxide-triggered cGMP generation in BM MKs. In summary, we established for the first-time optogenetics in primary MKs and show that PDE5 is the predominant PDE regulating cGMP levels in MKs. These findings also demonstrate that optogenetics allows for the precise manipulation of MK biology.     

高山峡谷地区无线电遥测与GPS 空间定位的比较:野外放归大熊猫的跟踪定位

以佩戴具有无线电发射功能的GPS 颈圈(Lotek GPS_ 4400M) 的放归大熊猫“祥祥”作为目标动物, 2006 年4 月至2007 年2 月,采用无线电遥测技术(RT)和GPS 跟踪技术在卧龙自然保护区的“五一棚”区域, 每日监测大熊猫在野外环境下的生存状况、移动规律和觅食行为。为了比较RT 和GPS 在高山峡谷地区空间定位的可行性和有效性,我们引入空间定位率、地形特征、空间定位差、巢域大小和日移动距离等指标来分析RT 和GPS 之间的定位差异。结果表明:RT 的空间定位效率明显高于GPS 的自动定位(P < 0. 001),分别是54.1%(绘图法)和45. 2% (≧ 2D);不同月份RT 和GPS 的空间定位率之间具有显著性差异(P < 0. 05),这与大熊猫不同月份的海拔活动范围和觅食行为特性密切相关。RT 位点的地形指数中坡度高于GPS,坡向和海拔高度较GPS 定位点低,两种无线电遥测方法(两点直接计算法和绘图法)之间没有显著性差异(P > 0.05);同一天位点之间的距离(空间定位差)平均450 ～ 660 m 左右;RT 与GPS 所估测的大熊猫巢域大小,除5 月、9 月和12月RT 低于GPS 外,其余月份为前者高于后者,但无显著性差异(P > 0. 05);日移动距离除12 月份RT 小于GPS 外,其余月份都呈现出RT 大于GPS 的格局,统计检验结果两者之间差异显著(P < 0. 05);两种无线电遥测方法所测指数之间都无明显差异(P > 0. 05)。这说明RT 遥测和GPS 定位都可以应用于高山峡谷地区野生动物的生态学研究,而且GPS 无线电颈圈在亚高山和高山森林中具有可行性和有效性。     

Development and Characterization of a New TILLING Population of Common Bread Wheat (Triticum aestivum L.)

Mutagenesis is an important tool in crop improvement. However, the hexaploid genome of wheat (Triticum aestivum L.) presents problems in identifying desirable genetic changes based on phenotypic screening due to gene redundancy. TILLING (Targeting Induced Local Lesions IN Genomes), a powerful reverse genetic strategy that allows the detection of induced point mutations in individuals of the mutagenized populations, can address the major challenge of linking sequence information to the biological function of genes and can also identify novel variation for crop breeding. Wheat is especially well-suited for TILLING due to the high mutation densities tolerated by polyploids. However, only a few wheat TILLING populations are currently available in the world, which is far from satisfying the requirement of researchers and breeders in different growing environments. In addition, current TILLING screening protocols require costly fluorescence detection systems, limiting their use, especially in developing countries. We developed a new TILLING resource comprising 2610 M(2) mutants in a common wheat cultivar 'Jinmai 47'. Numerous phenotypes with altered morphological and agronomic traits were observed from the M(2) and M(3) lines in the field. To simplify the procedure and decrease costs, we use unlabeled primers and either non-denaturing polyacrylamide gels or agarose gels for mutation detection. The value of this new resource was tested using PCR with RAPD and Intron-spliced junction (ISJ) primers, and also TILLING in three selected candidate genes, in 300 and 512 mutant lines, revealing high mutation densities of 1/34 kb by RAPD/ISJ analysis and 1/47 kb by TILLING. In total, 31 novel alleles were identified in the 3 targeted genes and confirmed by sequencing. The results indicate that this mutant population represents a useful resource for the wheat research community. We hope that the use of this reverse genetics resource will provide novel allelic diversity for wheat improvement and functional genomics.     

发光细菌JMU07的分离鉴定及其在生物毒性测定的初步研究

从乌贼表皮通过分离纯化得到一株发光细菌JMU07。该菌的菌落形态呈典型细菌菌落特征;显微镜下观察其为球杆状菌,革兰氏染色阴性。用荧光分光光度计测定其发光波长在420-650 nm之间,最大发光波长为477 nm。16S rDNA法测序,构建系统进化树,初步鉴定发光细菌JMU07为鳆发光杆菌(Photobacterium leiog-nathi)。生长发光曲线测定表明,发光细菌JMU07发光强度最高出现在对数中后期,相比明亮发光杆菌,JMU07具有发光强度高,持续发光时间长的特点。根据国标GB/T15441-1995研究HgCl2浓度与发光细菌JMU07发光强度抑制率的关系得到:HgCl2浓度与发光细菌JMU07发光强度抑制呈良好线性关系;JMU07 EC50为0.11 mg/L,略低于明亮发光杆菌的0.14 mg/L,表明JMU07对HgCl2的毒性更敏感。因此新分离得到的发光细菌JMU07有希望用于环境检测、食品卫生与安全等领域综合毒性的快速检测。