玉米新老品种生理生态特征与竞争力差异研究

本文以中国不同年代主要的玉米品种为试验材料，深入研究了玉米品种更替过程中新老品种的生理生态特征和竞争力差异，分析了差异形成的原因并进行了理论探讨。 玉米新老品种竞争力差异的研究采用了单作和混作两种方式，设高低两个密度。生长过程中全面测量了生物量、形态、生理和群体指标，运用了生长分析的方法来研究竞争，整合了各水平参数来解释竞争结果，并用本研究数据检验了生长冗余理论。 玉米新老品种对比研究发现新品种的生理生态特征普遍优于老品种。这些优势不仅体现在较高的生物量积累、较大的籽粒库容和较强的再分配能力上，而且体现在高的叶面积指数、衰老过程中仍维持较高的叶绿素含量、可溶性蛋白含量和光合效率上，同时新品种的群体特性还具有更低的感病率和更少的无效分蘖。玉米新老品种竞争结果表明在混作条件下，相对总产量这一指标反映出新老品种间明显的互利效应，且这种效应随发育阶段而降低。新品种对老品种的相对竞争力则随着发育阶段波动，并且密度和发育阶段两因子对品种竞争力的影响有明显的相互作用。相对于新品种，老品种的确存在叶片和根系的生长冗余部分，但老品种并没有在混作竞争中获得明显的竞争优势，即玉米品种选育并不完全符合生长冗余理论，因此在理解植物竞争力方面仍需要其他层面更深入的探讨。 同时，玉米品种选育不完全符合生长冗余理论的结论在农学实践上并非毫无价值。因为新品种总生物量的提高，不仅增加了籽粒产量，而且增加了秸秆产量，提供了更多可利用的生物质资源。相对于仅仅关注粮食产量，综合利用中国农村巨大的生物质资源具有更重要的生态意义。

转基因油菜和野芥菜杂交种的适合度

转基因植物与野生亲缘种间的基因流动是目前生物安全的研究热点。甘蓝型油菜（Brassica napus 以下简称油菜）及其亲缘种是基因流研究中的模式植物体系，油菜的亲缘种之一野芥菜(B. juncea)在中国分布广泛，是常见的农田杂草。本文以转基因抗虫油菜和野芥菜为研究材料，分别用综合适合度(基于生长和生殖)和适合度(仅基于种子产量)的指标来评估其杂交种在模拟农田情况下的适合度，揭示转基因在自然环境中的命运，以期为中国的油菜基因流的管理和控制提供参考。主要结果如下： 由于杂种优势和亲本效应的影响，转基因杂交种在不同的种植季节的生长与长势较好的亲本相近。因而转基因杂交种的综合适合度介于双亲之间，或与表现较好的亲本相似。因结实较差，从种子产量上来看，转基因杂交种的适合度较低。杂交种的后代因母本效应保留了一定的休眠特性，这有助于杂交种在种子库中持续存在。野芥菜和杂交种秋播时在冬季覆大棚条件下的适合度较高，在全球变暖的情况下，野芥菜和杂交种有可能在原先较冷不适宜的地区生长，从而促进转基因的散布。 杂交实验表明，转基因花粉与野芥菜的亲和性略高于非转基因花粉，但不显著；在两种花粉发生竞争时，转基因花粉的亲和性显著较高。混合花粉授粉的结实率高于单种花粉的结实率。可见在转基因花粉在授粉阶段不存在适合度代价。同时两季秋播田间实验表明，转基因油菜与非转基因油菜的适合度没有明显差异，即在植株的生长结实阶段，当没有竞争和虫害选择压力时，转基因没有适合度代价。也证明转基因散布的潜在可能性较大。此外，转基因杂交种的开花量很大，而回交实验中转基因杂交种花粉的育性高于胚珠的育性。转基因有可能通过持续的回交逃逸。 油菜的种子较大，野菜芥的种子较小，转基因油菜与野芥菜的杂交种基本都是小种子，其种子直径低于双亲。种子大小影响了发芽率、出苗率和营养生长期。小种子出苗率和发芽率较低，营养生长期较长。在没有竞争的条件下，春播实验中，种子大小对植物的适合度及综合适合度没有影响。秋播实验中种子大小对除转基因油菜外的所有植物基因型的综合适合度有显著影响。但对于依据种子产量计算的适合度指标来说，种子大小仅对野芥菜有影响，而对其它基因型影响不大。竞争扩大了种子大小之间生长表现的差异。竞争条件下，不同种子级别间的生长变异系数大于无竞争的情况。 总之，转基因花粉较强的竞争能力和其与野芥菜的亲和性使转基因杂交种的产生成为可能。来自母本的休眠特性有助于杂种种子在种子库中存在并在条件适宜时萌发、开花，有助于转基因的进一步扩散。转基因杂交种具有较高的综合适合度，虽然结实率很低，但其开花量比较大，可能会产生大量可育的花粉，在与野生亲本的持续回交过程中，转基因有可能逃逸。同时，转基因杂交种的种子级别偏小有可能会加剧这种逃逸。

超高产杂交稻Oryza sativa L.光合作用和光抑制特性的研究

较系统地比较研究了超高产杂交稻两优培九培矮64S 93-11和华安3号X07S 紫恢100与多年来大规模推广种植的杂交稻品种汕优63珍汕97A 明恢63的光合生理特性结果表明 1 从苗期到抽穗期超高产杂交稻两优培九和华安3号的净光合速率Pn都比汕优63高而在苗期的午间强光条件下和分蘖期的早晨以及抽穗期的早晚相对弱光条件下其Pn的差别尤为显著说明超高产杂交稻两优培九和华安3号不仅有较高的Pn和较强的抗光抑制能力而且还能充分利用早晨和傍晚较弱的光照条件有效地进行光合作用 2 超高产杂交稻剑叶具有较高的光合色素含量和Chla/b比值同时也具有较高的水分利用效率WUE较高的Chla/b比值表明超高产杂交稻剑叶能够更有效地利用太阳能而较高的WUE则有利于后期节约稻田用水 3 两优培九和华安3号类囊体膜的77K荧光光谱在不同发育时期均高于对照汕优63对其进行Gaussan分析发现这两个超高产杂交稻的反应中心以及天线复合物的发射峰均优于汕优63表明超高产杂交稻具有更强光能吸收能力并且能够将所吸收的光能高效地应用于光合电子传递 4 超高产杂交稻在苗期和分蘖期的净光合速率Pn都明显高于对照可以为群体的扩大后期的生长发育和高产奠定坚实的物质基础 5 三个杂交稻品种抽穗期剑叶的净光合速率相差不大但两优培九和华安3号具有较汕优63高得多的表观量子效率和羧化效率即超高产杂交稻能够高效地利用光能和田间二氧化碳首次提出对光能和二氧化碳的高效利用是两优培九和华安3号高产地重要原因 在对杂交稻净光合速率日变化的研究中发现超高产杂交稻两优培九和华安3号在午间强光条件下具有较对照汕优63更高的净光合速率表明超高产杂交稻具有更强的抗光抑制能力为了研究其光保护机理进一步研究了杂交稻对光抑制的响应结果表明 1超高产杂交稻两优培九和华安3号较对照品种汕优63具有更强的抗光抑制及光保护能力同时在光抑制结束后又能够更迅速地恢复光合功能较强的抗光抑制能力和较高的恢复能力可能是其高产的重要生理原因之一 2光抑制过程中超高产杂交稻叶黄素循环玉米黄素积累速率和积累量都明显高于对照并且在其后的恢复过程中其恢复速率和恢复程度也明显高于汕优63发现叶黄素循环的脱环化作用在光抑制处理30min时即基本接近最大值并未随着光抑制的进一步加重而不断上升认为叶黄素循环在杂交稻光保护中的重要作用可能在于玉米黄素的快速积累对光保护作用的启动 3在对自然条件下光抑制的研究中发现汕优63比超高产杂交稻两优培九和华安3号更容易受到午间光抑制的伤害 4午间光抑制条件下叶黄素循环的玉米黄素Z和环氧玉米黄素A大量积累而叶黄素循环库则没有什么变化认为是叶黄素循环脱环化组分A和Z的积累而不是叶黄素循环库对水稻在中午强光条件下的光保护起重要作用 5在所研究水稻品种的午间光抑制实验中叶绿素荧光的非光化学猝灭系数和叶黄素循环的脱环化状态DES之间没有正比例关系进一步推论环式电子传递可能在杂交稻的光保护中起重要作用 6对用不同试剂处理的杂交稻叶片进行光抑制处理研究发现ASAVDE酶底物其含量可以有效地调节活性处理对杂交稻的抗光抑制能力并没有带来多大改善而DTTVDE酶的特异抑制剂处理也没有使其光抑制大大加重而用DBMIB环式电子传递抑制剂处理则使杂交稻受到比对照强得多的光抑制对qN解析的结果发现强光下qE并未上升反而下降而qT却在光抑制条件下表现出上升现象这些实验结果首次阐明叶黄素循环的热耗散在杂交稻的光保护中不起关键作用而环式电子传递则对于杂交稻的光保护起至关重要的作用其机理可能在于强光条件下环式磷酸化的加剧生成大量ATP用于光破坏的修复作用同时避免类囊体膜的过度酸化从而导致强光下qN的下降这也是光抑制条件下qE下降和qT上升的原因所在此外在研究中发现光抑制处理导致Chla/b比值的上升并且提出这种上升的原因可能在于强光条件下光合系统对LHCII需求减少从而导致对Chlb需求减少最终使得部分Chlb向Chla转化这种转化可能是杂交稻在光抑制条件下的一种保护性响应 7对超高产杂交稻华安3号冠层不同衰老程度叶片的光合功能比较研究的结果表明剑叶的光合功能最强第二叶次之第三叶具有一定的光合功能第四和第五叶则相当衰老基本上丧失光合能力而光合机构的衰老可能始于反应中心的衰老天线系统的衰老要迟于反应中心的衰老叶片衰老进程中Chla和Chlb同步降解但是Chlb先还原为Chla导致Chla/b比值的上升并且认为衰老过程中的这种Chlb的还原是Chlb降解的一个早期的和不可避免的步骤

Assessment of cattle genetic introgression into domestic yak populations using mitochondrial and microsatellite DNA markers

Hybridization between yak Poephagus grunniens and taurine Bos taurus or indicine B. indicus cattle has been widely practiced throughout the yak geographical range, and gene flow is expected to have occurred between these species. To assess the impact of cattle admixture on domestic yak, we examined 1076 domestic yak from 29 populations collected in China, Bhutan, Nepal, India, Pakistan, Kyrgyzstan, Mongolia and Russia using mitochondrial DNA and 17 autosomal microsatellite loci. A cattle diagnostic marker-based analysis reveals cattle-specific mtDNA and/or autosomal microsatellite allele introgression in 127 yak individuals from 22 populations. The mean level of cattle admixture across the populations, calculated using allelic information at 17 autosomal microsatellite loci, remains relatively low (mY(cattle) = 2.66 +/- 0.53% and Q(cattle) = 0.69 +/- 2.58%), although it varies a lot across populations as well as among individuals within population. Although the level of cattle admixture shows a clear geographical structure, with higher levels of admixture in the Qinghai-Tibetan Plateau and Mongolian and Russian regions, and lower levels in the Himalayan and Pamir Plateau region, our results indicate that the level of cattle admixture is not significantly correlated with the altitude across geographical regions as well as within geographical region. Although yak-cattle hybridization is primarily driven to produce F-1 hybrids, our results show that the subsequent gene flow between yak and cattle took place and has affected contemporary genetic make-up of domestic yak. To protect yak genetic integrity, hybridization between yak and cattle should be tightly controlled.

Variety of antimicrobial peptides in the Bombina maxima toad and evidence of their rapid diversification

Antimicrobial peptides secreted by the skin of many amphibians play an important role in innate immunity. From two skin cDNA libraries of two individuals of the Chinese red belly toad (Bombina maxima), we identified 56 different antimicrobial peptide cDNA sequences, each of which encodes a precursor peptide that can give rise to two kinds of antimicrobial peptides, maximin and maximin H. Among these cDNA, we found that the mean number of nucleotide substitution per non-synonymous site in both the maximin and maximin H domains significantly exceed the mean number of nucleotide substitution per synonymous site, whereas the same pattern was not observed in other structural regions, such as the signal and propiece peptide regions, suggesting that these antimicrobial peptide genes have been experiencing rapid diversification driven by Darwinian selection. We cloned and sequenced seven genes amplified from skin or liver genomic DNA. These genes have three exons and share the same gene structure, in which both maximin and maximin H are encoded by the third exon. This suggests that alternative splicing and somatic recombination are less likely to play a role in creating the diversity of maximins and maximin Hs. The gene trees based on different domain regions revealed that domain shuffling or gene conversion among these genes might have happened frequently.

Differential expression of the Brunol/CELF family genes during Xenopus laevis early development

The BRUNOL/CELF family of RNA-binding proteins plays important roles in post-transcriptional regulation and has been implicated in several developmental processes. In this study, we describe the cloning and expression patterns of five Brunol genes in Xenopus laevis. Among them, only Brunol2 is maternally expressed and the zygotic expression of the other four Brunol genes starts at different developmental stages. During Xenopus development, Brunol1, 4-5 are exclusively expressed in the nervous system including domains in the brain, spinal cord, optic and otic vesicles. Brunol2 and 3 are expressed in both the somatic mesoderm and the nervous system. Brunol2 is also extensively expressed in the lens. In transfected Hela cells, BRUNOL1, 2 and 3 proteins are localized in both the cytoplasm and the nucleus, while BRUNOL4 and 5 are only present in the cytoplasm, indicating their different functions.

Microsatellite DNA analysis proves nucleus of interspecies reconstructed blastocyst coming from that of donor giant panda

A method for DNA isolation from early development of blastocyst and further analysis of nuclear and mitochondrial DNA was developed in present study. Total DNA was prepared from interspecies reconstructed blastocyst and a giant panda specific microsatellite locus g(010) was successfully amplified. DNA sequencing of the PCR product showed that two sequences of reconstructed blastocysts are the same as that of positive control giant panda. Our results prove that the nucleus of interspecies reconstructed blastocyst comes from somatic nucleus of donor giant panda.

Interspecies implantation and mitochondria fate of panda-rabbit cloned embryos

Somatic cell nuclei of giant pandas can dedifferentiate in enucleated rabbit ooplasm, and the reconstructed eggs can develop to blastocysts. In order to observe whether these interspecies cloned embryos can implant in the uterus of an animal other than th

Phantom mutation hotspots in human mitochondrial DNA

Phantom mutations are systematic artifacts generated in the course of the sequencing process. Contra common belief these artificial mutations are nearly ubiquitous in sequencing results, albeit at frequencies that may vary dramatically. The amount of artifacts depends not only on the sort of automated sequencer and sequencing chemistry employed, but also on other lab-specific factors. An experimental study executed on four samples under various combinations of sequencing conditions revealed a number of phantom mutations occurring at the same sites of mitochondrial DNA (mtDNA) repeatedly. To confirm these and identify further hotspots for artifacts, > 5000 mtDNA electropherograms were screened for artificial patterns. Further, > 30000 published hypervariable segment 1 sequences were compared at potential hotspots for phantom mutations, especially for variation at positions 16085 and 16197. Resequencing of several samples confirmed the artificial nature of these and other polymorphisms in the original publications. Single-strand sequencing, as typically executed in medical and anthropological studies, is thus highly vulnerable to this kind of artifacts. In particular, phantom mutation hotspots could easily lead to misidentification of somatic mutations and to misinterpretations in all kinds of clinical mtDNA studies.

Epigenetic marks in cloned rhesus monkey embryos: Comparison with counterparts produced in vitro

Until now, no primate animals have been successfully cloned to birth with somatic cell nuclear transfer (SCNT) procedures, and little is known about the molecular events that occurred in the reconstructed embryos during preimplantation development. In man

HSPC117 deficiency in cloned embryos causes placental abnormality and fetal death

Somatic cell nuclear transfer (SCNT) has been successfully used in many species to produce live cloned offspring, albeit with low efficiency. The low frequency of successful development has usually been ascribed to incomplete or inappropriate reprogramming of the transferred nuclear genome. Elucidating the genetic differences between normal fertilized and cloned embryos is key to understand the low efficiency of SCNT. Here, we show that expression of HSPC117, which encodes a hypothetical protein of unknown function, was absent or very low in cloned mouse blastocysts. To investigate the role of HSPC117 in embryo development, we knocked-down this gene in normal fertilized embryos using RNA interference. We assessed the post-implantation survival of HSPC117 knock-down embryos at 3 stages: E9 (prior to placenta formation); E12 (after the placenta was fully functional) and E19 (post-natal). Our results show that, although siRNA-treated in vivo fertilized/produced (IVP) embryos could develop to the blastocyst stage and implanted without any difference from control embryos, the knock-down embryos showed substantial fetal death, accompanied by placental blood clotting, at E12. Furthermore, comparison of HSPC117 expression in placentas of nuclear transfer (NT), intracytoplasmic sperm injection (ICSI) and IVP embryos confirmed that HSPC117 deficiency correlates well with failures in embryo development: all NT embryos with a fetus, as well as IVP and ICSI embryos, had normal placental HSPC117 expression while those NT embryos showing reduced or no expression of HSPC117 failed to form a fetus. In conclusion, we show that HSPC117 is an important gene for post-implantation development of embryos, and that HSPC117 deficiency leads to fetal abnormalities after implantation, especially following placental formation. We suggest that defects in HSPC117 expression may be an important contributing factor to loss of cloned NT embryos in vivo. (C) 2010 Elsevier Inc. All rights reserved.

A critical reassessment of the role of mitochondria in tumorigenesis

Background Mitochondrial DNA (mtDNA) is being analyzed by an increasing number of laboratories in order to investigate its potential role as an active marker of tumorigenesis in various types of cancer. Here we question the conclusions drawn in most of these investigations, especially those published in high-rank cancer research journals, under the evidence that a significant number of these medical mtDNA studies are based on obviously flawed sequencing results. Methods and Findings In our analyses, we take a phylogenetic approach and employ thorough database searches, which together have proven successful for detecting erroneous sequences in the fields of human population genetics and forensics. Apart from conceptual problems concerning the interpretation of mtDNA variation in tumorigenesis, in most cases, blocks of seemingly somatic mutations clearly point to contamination or sample mix-up and, therefore, have nothing to do with tumorigenesis. Conclusion The role of mitochondria in tumorigenesis remains unclarified. Our findings of laboratory errors in many contributions would represent only the tip of the iceberg since most published studies do not provide the raw sequence data for inspection, thus hindering a posteriori evaluation of the results. There is no precedent for such a concatenation of errors and misconceptions affecting a whole subfield of medical research.

Detection of hybridization between two loach species (Paramisgurnus dabryanus and Misgurnus anguillicaudatus) in wild populations

Artificial interspecific hybrids between large scale loach P. dabryanus and tetraploid pond loach M. anguillicaudatus (Cobitidae, Cypriniformes) are viable. To detect the occurrence of possible natural hybridization, genetic analyses by using microsatellite markers were performed for natural populations of large scale loach and pond loach, the reciprocal laboratory hybrids, and "supposed hybrids" with ambiguous morphology. The fertility of the artificial hybrids was also tested. At one diagnostic microsatellite (Mac50), one out of 20 "supposed hybrids" was identified to be F-1 hybrid between the two loach species because it had the same genotype as that of the laboratory hybrids. The triploid hybrids between the two species were confirmed to be female-sterile. The results show that rare hybridization has occurred between diploid large scale loach and tetraploid pond loach in nature although it may have little effect in genetic introgression. This study is helpful for fish conservation and encourages further investigation on natural hybridization and introgression of loaches.

Expression pattern of cellular nucleic acid-binding protein (CNBP) during embryogenesis and spermatogenesis of gibel carp

By differential screening, we cloned the CagCNBP, demonstrated its predominant expression in ovary and testis, and reported its development behavior during folliculogenesis and oogenesis by immunofluorescence localization (Liu and Gui, Gene 365:181-192, 2005), but its developmental behavior during spermatogenesis and its transcript distribution during embryogenesis are not revealed. In the present study, by in situ hybridization, we analyze CagCNBP expression pattern during gibel carp embryogenesis. The CagCNBP transcripts ubiquitously distributed in all embryonic cells in early developmental stage embryos, and peak in midbrain, hindbrain and somites of gibel carp larva during organogenesis. By antibody detection, we reveal CagCNBP protein distribution change during spermatogenesis. The cell-specific distribution of CagCNBP is revealed by immunofluorescence staining, and predominant CagCNBP expression in testis somatic cells and spermatogonia is demonstrated in this paper. For the first time, the CNBP distribution during spermatogenesis in vertebrate has been revealed.