Light-responsive cryptochromes from a simple multicellular animal, the coral Acropora millepora

Hundreds of species of reef-building corals spawn synchronously over a few nights each year, and moonlight regulates this spawning event. However, the molecular elements underpinning the detection of moonlight remain unknown. Here we report the presence of an ancient family of blue-light-sensing photoreceptors, cryptochromes, in the reef-building coral Acropora millepora. In addition to being cryptochrome genes from one of the earliest-diverging eumetazoan phyla, cry1 and cry2 were expressed preferentially in light. Consistent with potential roles in the synchronization of fundamentally important behaviors such as mass spawning, cry2 expression increased on full moon nights versus new moon nights. Our results demonstrate phylogenetically broad roles of these ancient circadian clock-related molecules in the animal kingdom.     

Mass Spawning by Green Algae on Coral Reefs

Predawn episodes of mass spawning by green algae (up to nine species in five genera on a single morning) intermittently cloud Caribbean waters. Species- and sex-specific bouts of anisogamous gamete release occurred synchronously and predictably on a given morning, with closely related species spawning at different times. Algal sexual reproduction was seasonal, but, unlike the mass-spawning behavior of other sessile marine organisms, showed no lunar or tidal cycling. The discovery of mass-spawning behavior by these algae has important implications for future studies of the reproductive ecology and speciation of a vital, yet poorly understood, component of the coral reef community.     

绿盲蝽卵黄原蛋白(Vg)基因片段克隆及不同生境表达量分析

为了探讨不同果园生境对绿盲蝽生殖的影响,从分子角度确定绿盲蝽适合的产卵生境,本研究首先采用绿盲蝽专用诱芯调查了枣园、葡萄园的绿盲蝽发生量,然后通过比较半翅目,直翅目,膜翅目,鳞翅目等昆虫cDNA同源序列,根据基因保守序列设计一对简并性引物,克隆绿盲蝽卵黄原蛋白(Vg)基因片段,采用RT-PCR的方法检测了2种生境中Vg基因相对表达量。结果显示,枣园绿盲蝽的发生量始终高于葡萄园,枣园有4个发生高峰,而葡萄园有2个。基因克隆获得一段大小为732 bp的基因片段,编码244个氨基酸;该基因序列与其他蝽类昆虫Vg基因氨基酸序列的同源性达80%以上;系统进化分析表明,绿盲蝽Vg基因与赤须盲蝽、烟盲蝽及黑肩绿盲蝽的亲缘关系最近,克隆的基因片段为绿盲蝽Vg基因片段,并命名为AL-Vg。不同生境Vg基因表达量比较可知,枣园的表达量低于葡萄园,结合发生量表明枣园生境更适合绿盲蝽生殖产卵。     

Mass spawning in tropical reef corals

Synchronous multispecific spawning by a total of 32 coral species occurred a few nights after late spring full moons in 1981 and 1982 at three locations on the Great Barrier Reef, Australia. The data invalidate the generalization that most corals have internally fertilized, brooded planula larvae. In every species observed, gametes were released; external fertilization and development then followed. The developmental rates of externally fertilized eggs and longevities of planulae indicate that planulae may be dispersed between reefs.     

Coral community reproductive patterns: red sea versus the great barrier reef

In contrast to many corals of the Great Barrier Reef, Australia, which are synchronous multispecific spawners, the abundant coral species in the northern Red Sea, Israel, exhibit temporal reproductive isolation. Spawning dates of 12 of the 13 Red Sea species followed lunar periodicity and were consistent throughout 3 years of study. Spawning periods of all species occurred in different seasons, different months, or different lunar phases within the same month. The high abundance of the corals studied at Eilat may be due in part to the advantages gained through not having overlapping spawning periods and settlement times.     

Hybridization and the evolution of reef coral diversity

Hundreds of coral species coexist sympatrically on reefs, reproducing in mass-spawning events where hybridization appears common. In the Caribbean, DNA sequence data from all three sympatric Acropora corals show that mass spawning does not erode species barriers. Species A. cervicornis and A. palmata are distinct at two nuclear loci or share ancestral alleles. Morphotypes historically given the name Acropora prolifera are entirely F(1) hybrids of these two species, showing morphologies that depend on which species provides the egg for hybridization. Although selection limits the evolutionary potential of hybrids, F(1) individuals can reproduce asexually and form long-lived, potentially immortal hybrids with unique morphologies.     

牛体外发育胚胎特定阶段差异表达基因研究

[目的]研究不同发育时期牛体外受精胚胎在基因表达模式上的差异。[方法]利用单个胚胎mRNA差异显示技术,对单个8细胞期胚胎与囊胚进行mRNA差异显示,获得1条特异表达条带,对其进行克隆、测序,并与GenBank进行对比。[结果]该序列与牛核糖体蛋白L31（ribosomal protein L31,RPL31）基因序列具有99%的同源性。采用实时定量PCR技术检测8细胞期和囊胚期胚胎RPL31的mRNA表达量,结果表明,RPL31在8细胞期胚胎的相对表达量为囊胚期胚胎的3.2倍。[结论]为揭示和阐明控制牛早期胚胎发育的相关机理提供依据。     

Climate change,human impacts,and the resilience of coral reefs

The diversity, frequency, and scale of human impacts on coral reefs are increasing to the extent that reefs are threatened globally. Projected increases in carbon dioxide and temperature over the next 50 years exceed the conditions under which coral reefs have flourished over the past half-million years. However, reefs will change rather than disappear entirely, with some species already showing far greater tolerance to climate change and coral bleaching than others. International integration of management strategies that support reef resilience need to be vigorously implemented, and complemented by strong policy decisions to reduce the rate of global warming.     

低能氩离子束介导将绿色荧光蛋白基因导入小麦的研究

用低能氩离子束介导将改良的绿色荧光蛋白基因（ｍＧＦＰ４）导入小麦栽培品种皖９２１０和皖麦３２号的成熟胚细胞。在含有１００￣１４０ｍ／Ｌ巴龙霉素培养基上继代培养后,获得了一批抗性愈伤组织。经过分化培养,皖９２１０获得５株再生苗,皖麦３２号获得３２株绿苗,对照的２００枚成熟胚未获得再生苗。对再生苗进行ＰＣＲ检测,均扩增出６００ｂｐ的ｎｐｔⅡ基因片段。取４株长势好的绿苗进行Ｓｏｕｔｈｅｒｎ杂交分析,结果     

Siphonodictidine, a Metabolite of the Burrowing Sponge Siphonodictyon sp. That Inhibits Coral Growth

Siphonodictidine is the major secondary metabolite of an undescribed Indo-Pacific sponge Siphonodictyon sp. that burrows into living coral heads. The structure of siphonodictidine was determined from spectral data. Laboratory bioassays suggest that siphonodictidine and, by analogy, the siphonodictyals from S. coralliphagum are responsible for maintaining zones of dead coral polyps around the oscular chimneys of these sponges.     

体外培养山羊早期胚胎基因表达的研究

用单胚构建的mRNA差异显示技术,对体外培养的山羊早期2、4、8～16细胞期胚胎的基因表达进行研究,并选择1条在8～16细胞期胚胎特异表达的条带进行分析。结果表明：该片段与牛NADH脱氢酶1a亚复合体4基因具有97％的同源性。该基因影响细胞内ATP的合成,可能与羊早期体外培养胚胎的正常发育有关。     

安康地区泥鳅胚胎发育研究

[目的]了解安康地区泥鳅的胚胎发育全过程。[方法]用DOM和LHRH-A2对泥鳅进行催产,将所产卵子进行人工授精后所获的胚胎置于24～26℃、pH值7.0～7.5的淡水中培养,进行泥鳅的胚胎发育观察,详细描述从受精卵到初孵仔鱼期的27个发育时期的形态特征和发育时间。[结果]泥鳅胚胎发育可划分为卵裂期、囊胚期、原肠胚期、神经胚期和器官形成期。在24～26℃、pH值7.0～7.5的淡水中,泥鳅胚胎从受精到孵化出膜历时30h45min。[结论]为泥鳅的人工繁殖和遗传育种研究提供了重要的参考。     

瘤背石磺产卵规律与天文潮汐关系

为了开展瘤背石磺人工增养殖技术以满足日益增长的市场需求,弄清瘤背石磺的产卵规律是至关重要的。该实验比较三年来瘤背石磺产卵规律趋势,以及潮汐潮高与产卵周期关系,从2013年至2015年,在每年的7月到9月间连续购进同地区的野生瘤背石磺,选取同等规格的瘤背石磺暂养后记录每批瘤背石磺产卵情况,并与潮汐规律以及潮位高度相比较。结果显示:按农历计算,瘤背石磺在每个月的小潮(初七、二十三)前后集中大量产卵,维持6~7 d,在大潮(初一、十五)来临之前一至两天结束产卵,趁大潮时的潮水运动将已出膜的幼体带入海洋。在海水中经过15 d左右的发育,待瘤背石磺发育至大面盘幼虫期时再随下一次大潮回到滩涂。根据结果可以得出以下结论:(1)瘤背石磺的产卵规律与天文潮汐相关,呈现出伴随着小潮出现一次产卵高峰,并且会在开始产卵后的两至三天内出现峰值;(2)理论上说,从小潮到下一个小潮为14 d 19 h55 min,实际上,由于潮汐的迟滞性等原因,瘤背石磺的产卵周期为14 d 21 h 20 min。     

转录靶向猪Jiv基因shRNA细胞株的建立 及抗猪瘟病毒转基因猪的构建

【目的】构建转入靶向猪Jiv基因shRNA干扰片段的阳性细胞株,通过比较各细胞株对猪瘟病毒增殖的干扰效果,筛选对猪瘟病毒增殖有明显抑制作用的细胞株,为抗猪瘟转基因猪的构建提供材料。【方法】研究设计了靶向猪Jiv基因的4个shRNA干扰片段,并构建插入干扰片段的慢病毒(P1、P2、P3、P4)。将慢病毒分别转染PK-15细胞,阳性细胞接种猪瘟病毒后72 h用实时荧光定量PCR检测猪瘟病毒RNA的量,以比较4种干扰细胞株对猪瘟病毒增殖的干扰效果。将对猪瘟病毒增殖有较好干扰效果的P2慢病毒干扰载体转染猪胎儿成纤维细胞,获得稳定表达靶向猪Jiv基因shRNA干扰片段的猪胎儿成纤维细胞株,作为抗猪瘟病毒转基因猪构建的供体细胞,将细胞核移植到成熟的去核猪卵母细胞中,获得体细胞核移植胚胎,移植入受体母猪输卵管中进行转基因猪构建,对获得的转基因猪进行外源基因的鉴定。【结果】获得了4株转入靶向猪Jiv基因shRNA干扰片段的PK-15细胞株,其中转入P2干扰载体的细胞株对猪瘟病毒的增殖有明显的抑制作用,将转入P2干扰载体的猪胎儿成纤维细胞株为核供体细胞通过体细胞核移植,获得经鉴定为外源基因插入阳性的转基因猪。【结论】细胞Jiv基因的表达对猪瘟病毒的增殖有一定的影响,筛选获得的一个干扰细胞株对猪瘟病毒的增殖有明显的干扰效果;通过体细胞核移植技术获得一头转入靶向猪Jiv基因shRNA干扰片段（P2）的转基因猪。     

未来气候变化情景下西北太平洋柔鱼资源补充量预测

结合1996—2005年西北太平洋柔鱼经验产卵场、推测产卵场和索饵场各月适宜水温面积占总面积的比例(P;),分析其与柔鱼单位捕捞努力量渔获量(catch per unite effort, CPUE)之间的相关关系,选取统计关系显著的P;与CPUE建立线性预报模型,在RCP4.5和RCP8.5两种情景下,对2025、2055和2095年柔鱼资源补充量进行预测。结果表明:在气候变化下,柔鱼经验产卵场适宜SST范围向北移动,到2095年移动至经验产卵场最北缘,1—4月经验产卵场平均P;呈下降趋势:但变化不显著;推测产卵场适宜SST范围向北移动,到2095年移动至推测产卵场最北缘,且已超过推测产卵场范围,1—4月推测产卵场平均P;呈下降趋势,变化显著;索饵场适宜SST范围向北移动且有扩张趋势,7—10月索饵场平均P;呈下降趋势,变化显著。相关分析表明,1996—2005年2月和3月推测产卵场P;与CPUE显著正相关。未来柔鱼CPUE呈下降趋势:到2025年,柔鱼CPUE为(208.87±5.46)t/艘;到2055年,为(198.00±47.92)t/艘;到2095年,为(154.35±48.72)t/艘。到2095年,相比于2000年柔鱼CPUE最大下降60.08%。建议采取兼容气候适应性的柔鱼资源养护管理措施。     

池养珠江斑鳠人工繁殖和胚胎发育的初步研究

对在养殖池塘中培育珠江斑鳠Mystus guttatus亲鱼和人工繁殖的主要方法进行了总结,并提出:斑鳠雌、雄亲鱼的外观特征可作为鉴别其性成熟度的依据;对斑鳠雌鱼进行人工催产时宜分2次注射(间隔时间为24h),第1针注射PG(鲤)+LRH-A+DOM,第2针注射LRH-A+DOM+HCG。对斑鳠进行人工授精后,在水温27.5℃的条件下,受精卵约经17h30m in进入原肠期,胚胎孵化时间约为55h。     

天蚕不同蛾色产卵与幼虫体色的调查

对不同蛾色分离、产卵数量、幼虫体色、茧色和茧质调查结果表明：蛾色不同其造卵数和产卵数有差异,产卵数量最高的是黄色蛾早×黄色蛾6组合,平均为260粒,产卵净度达86．5％。浅灰色、黑色和暗黑色产卵量较低;幼虫体色可分为浅黄色和绿色2个类型,幼虫体色不同,其茧色也有差异,可分为黄色、浅黄色、浅绿色和绿色;不同茧色的茧层率差异不显著,雄茧高于雌茧的茧层率1％左右。     

影响羊早期胚胎发育阻滞相关基因的筛选

采用单胚构建的mRNA差异显示技术,探讨经体内发育和体外培养获得的山羊8~16细胞期胚胎基因的表达情况,并从在体内发育胚胎特异表达的片段中选择1个片段进行分析。结果表明:该片段与牛肥胖基因具有86%的同源性。该基因对卵母细胞的成熟以及早期胚胎的分裂和分化具有明显的作用,可调节母体的妊娠生理,影响生殖激素的分泌,从而对克服早期胚胎发育阻滞具有重要的作用。     

克氏原螯虾不同生理时期肝胰腺和肌肉生化组成及能量的变化

研究分析了克氏原螯虾（Procambarus clarkii）雌性亲本在产卵和抱卵期间肝胰腺和肌肉生化组成的变化及能量变动。结果表明：（1）克氏原螯虾产卵前后肝胰腺与肌肉指数均无显著性变化（P>0.05）,而卵巢指数由6.14%降到0.40%（P<0.01）,产卵后至卵破膜阶段肝胰腺指数上升了13.29%,显著高于产卵前水平（P<0.05）。（2）克氏原螯虾产卵后肝胰腺和肌肉中3大营养物质均呈现下降趋势,其中肝胰腺中粗脂肪含量与肌肉中粗蛋白含量下降极显著（P<0.01）,至卵破膜阶段,肝胰腺中的蛋白质和碳水化合物含量快速恢复且极显著超过产卵前水平（P<0.01）。产卵前后肝胰腺能值下降最大,抱卵期间肌肉能值下降显著。（3）产卵后,多不饱和脂肪酸（PUFA）显著下降,C22∶6n-3(DHA)与C20∶5n-3(EPA)下降幅度最大,分别为44.33%,30.80%。在这3个阶段,C20∶4n-6(AA)一直呈上升趋势,饱和脂肪酸（SFA）和单不饱和脂肪酸（MUFA）较为稳定,略有上升。     

转绿色荧光蛋白标记基因猪整合检测的研究

[目的]为转基因猪生产的整合效率、外源目的基因表达水平、目的基因在猪体内组织特异性表达分布等相关领域的进一步研究奠定基础。[方法]用显微注射法将GFP标记的转基因猪表达载体PM1导入湖北白猪和通湖猪1-细胞、2-细胞胚胎中,胚胎移植妊娠后生产仔猪,取耳或尾组织样提取基因组总DNA,用PCR法检测PM1在猪基因组中的整合情况。[结果]在所有生产的78头仔猪中,有6头猪用PCR检测为转PM1基因阳性猪,这6头猪在3次PCR扩增反应中都得到650 bp目的扩增片段。其他样品没有得到目的扩增片段的为阴性猪。[结论]载体PM1可以用于携带外源基因进行转基因猪生产。