Electroporation of immature maize zygotic embryos and regeneration of transgenic plants

Using the pulse-discharging electroporation system HPES-3, we have transferred the neomycin phosphotransferase II (nptII) gene and -glucuronidase (gus) gene into mechanically-woulded immature zygotic embryo cells of an elite local maize cultivar Huanong Supersweet No. 42 and have produced transgenic maize plants. DNA hybridization and NPTII dot assay showed that the foreign genes were integrated into the genomes and expressed stably in the cells of the transgenic calluses and plants.     

水稻胚囊壁的形成与发育观察

通过透射电镜对水稻(Oryza sativa L.)功能大孢子形成开始至胚囊成熟期间胚囊壁的形成与发育进行观察,结果表明:水稻胚囊壁是在原有功能大孢子壁的基础上,通过与其周围退化珠心细胞留下的壁相叠合,使壁加厚。功能大孢子近合点端壁存在胞间连丝,其中个别胞间连丝可保留到八核胚囊。胚囊壁上内突最早于四核胚囊近珠孔端发生。八核胚囊形成后,内突的发育在胚囊不同的细胞中表现不同,其中以中央细胞最具特点,表现为先在中央细胞与珠心相接的近珠孔端和近合点端两个区域的胚囊壁上形成,以后近珠孔端胚囊壁上的内突大量增加,而近合点端的却增加不明显,中部胚囊壁上的内突出现的时间相对较晚。到胚囊成熟时,近珠孔端胚囊壁上内突的分布密度最大,中部次之,近合点端的最小,三个区域上内突的形态各异。反足细胞与珠心相接的胚囊壁上内突的形成时间较早,但以后的发育却相对缓慢,数量增加不明显。2个助细胞交界处胚囊壁上的丝状器在胚囊未明显膨大时已形成。卵细胞除在与助细胞交界处的壁外,其它部位不形成明显的内突结构。     

Pulse treatments of penicillin-G and streptomycin minimise internal infections and have post-treatment effects on the morphogenesis of ginseng root culture

This study was initiated to determine whether antibiotic pulse treatments (APT) could effectively eliminate internal infections of ginseng (Panax ginseng) root explants containing vascular tissue, and subsequently have post-treatment effects on changing explant behaviors in callus induction and organogenesis or somatic embryogenesis. For contamination control, a treatment of 40 min with an antibiotic solution consisting of 1000 mg/1 of penicillin-G and 1000 mg/1 of streptomycin immediately following Na-hypochlorite sterilisation significantly decreased contamination rate. Extending treatment time to 2–3 h further lowered the contamination rate to 30–40%. On the other hand, explants treated with antibiotics for 20 min or less were all contaminated. APT also had post-treatment effects; it delayed callus induction for 1–12 months depending on pulse duration and stimulated the generation of more hardand darker looking than fragile- and lighter looking callus. The induced callus proliferated at a depressed rate, increasing subculture intervals from 1 to several weeks, and not until after five subcultures did it fully recover. The regeneration ability of the recovered callus was also affected by APT: the regeneration of adventitious roots was promoted, whereas somatic embryos were not observed.Abbreviations APT Antibiotic pulse treatments - 2,4-D 2,4-dichlorophenoxyacetic acid     

Conditions of induction of secondary dormancy in apple after breaking of primary dormancy by anaerobiosis and low temperature

Dormant embryos of Pyrus malus L. cv. Golden delicious, isolated from the fruits at harvest time or after a few months storage at 10 to 15°C, were kept under anaerobic conditions in order to eliminate primary dormancy. Germination tests were then carried out at different temperatures, using three modes of culture depending on the nature of the contact between the embryo and the medium. In CM the distal part of the two cotyledons was immersed in the medium. In RM only the embryonic axis was immersed. In C/2M the embryo was placed flat on the medium, the radicle and the external surface of one cotyledon being in contact with it.
Results showed that primary dormancy was released progressively depending on the duration of the anaerobic treatment. After a treatment of 11 or 13 days the last symptoms of primary dormancy were only apparent when germination tests were carried out at high temperatures (26–30°C) or in CM mode of culture.
When the embryos were kept at 4°C during 3 months inside the fruits, subsequent germination was inhibited at high temperature and in CM mode of culture. When the embryos were kept under anaerobic conditions (7 days) after the chilling treatmem inside the fruits, germination was no longer inhibited. It is concluded that the inhibition of germination at high temperature and in CM mode of culture is due to the persistence of traces of primary dormancy. Therefore, these conditions do not seem to induce secondary dormancy in apple embryos.
After elimination of primary dormancy by anaerobiosis. only application of (±) abscisic acid (3.8 and 19 μM) inhibited germination. These results support the idea that ABA is an important factor in the induction of dormancy. However, the question remains whether this secondary embryo dormancy has the same characteristics as the original primary dormancy.     

虎斑乌贼的胚胎耗氧率

为了探究虎斑乌贼胚胎不同发育时期的耗氧率变化和几种生态因子对胚胎发育过程耗氧率的影响,试验采用封闭静水装置,对不同发育时期(12期)的耗氧率进行测定,并研究不同盐度(21、24、27、30、33)、温度(18、21、24、27、30 ℃)和pH(7.0、7.5、8.0、8.5、9.0)对胚胎4个主要发育时期(受精卵期、原肠胚期、器官形成期和内骨骼形成期)耗氧率的影响.结果表明: 胚胎各个发育时期耗氧率不同,随着发育的进程而增大,受精卵期为0.082 mg·(100 eggs)^-1·h^-1,而到原肠胚期的耗氧率显著升高,为0.279 mg·(100 eggs)^-1·h^-1,到孵化期时,耗氧率达到1.367 mg·(100 eggs)^-1·h^-1;盐度对器官形成期和内骨骼形成期的耗氧率均有显著影响(P<0.05),对受精卵期和原肠胚期影响不显著(P>0.05),当盐度为30时,4个发育时期耗氧率均达到最大值,分别为0.082、0.200、0.768和1.301 mg·(100 eggs)^-1·h^-1;温度对原肠胚期、器官形成期和内骨骼形成期的耗氧率有显著影响(P<0.05),对受精卵期无显著性影响(P>0.05),在27 ℃时,胚胎4个发育时期均达到最大值,分别为0.082、0.286、0.806和1.338 mg·(100 eggs)^-1·h^-1;而pH对4个发育时期的耗氧率均无显著性影响(P>0.05),受精卵期在pH 8.0时达到最大值,为0.116 mg·(100 eggs)^-1·h^-1,原肠胚期、器官形成期、内骨骼形成期在pH 8.5时达到最大值,分别为0.281 、0.799和1.130 mg·(100 eggs)^-1·h^-1.     

TLR4激活剂对约氏疟原虫感染早期免疫应答的影响

通过外源性给予LPS,探讨TLR4在疟疾感染早期免疫应答的作用特点及其免疫调节作用。通过Plasmodium yoelii 17XL感染的BALB/c小鼠建立鼠疟模型并在感染前给予LPS,于感染第0、3和5天制备脾细胞悬液,通过流式细胞术检测脾细胞悬液中TLR4+DCs和Tregs百分含量;ELISA方法检测脾细胞培养上清中IFN-γ和IL-10水平。结果显示,LPS处理能够显著延长宿主生存期,降低原虫血症水平,同时显著提升脾上清中的IFN-γ水平,降低抑炎性细胞因子IL-10水平。在感染早期,LPS处理可诱导Th1型免疫应答的有效建立,明显遏制P.y 17XL红内期疟原虫的感染进程。     

斑马鱼胚胎细胞和中国仓鼠卵巢细胞远缘杂交融合条件的优化

以斑马鱼胚胎细胞(ZEM-2s)和中国仓鼠卵巢细胞(CHO-k1)为实验材料,采用聚乙二醇(PEG)作为促融剂,从PEG的相对分子质量、浓度、作用温度和时间等方面进行单因子实验,以期寻找两种细胞融合的最佳条件。实验结果表明,融合的最适条件是融合温度为37℃,浓度为40%,分子量为2 000的PEG处理斑马鱼胚胎细胞和中国仓鼠卵巢细胞100sec,平均融合率高达25.3%,与未加入PEG的细胞相比,最佳条件下处理的两种细胞融合现象明显(p<0.05),表明该条件下的处理能够显著促进细胞的融合。     

Massacre of the innocents in Cardiocondyla thoracica: Manipulation by adult ant males incites workers to kill their immature rivals

Wingless males of the ant genus Cardiocondyla exhibit a remarkable range of intrasexual competition, from mutual tolerance among males in the colonies of some species to regular lethal fighting in the nests of others. Here, we describe male–male antagonism in Cardiocondyla thoracica (Smith, 1859) from Northern Australia. Males appear to widely ignore adult competitors, but grab defenseless, young rivals during or immediately after emergence and daub them with hindgut secretion. This fluid elicits fatal worker aggression against the besmeared individual. Besmearing and subsequent worker aggression is known from other species of Cardiocondyla but is usually directed towards adult contenders, whereas males kill freshly eclosing males by biting or dismembering. The behavior of C. thoracica males thus constitutes a novel nuance of rival elimination.     

Inbreeding and purging at the genomic Level: the Chillingham cattle reveal extensive,non‐random SNP heterozygosity

Local breeds of livestock are of conservation significance as components of global biodiversity and as reservoirs of genetic variation relevant to the future sustainability of agriculture. One such rare historic breed, the Chillingham cattle of northern England, has a 350‐year history of isolation and inbreeding yet shows no diminution of viability or fertility. The Chillingham cattle have not been subjected to selective breeding. It has been suggested previously that the herd has minimal genetic variation. In this study, high‐density SNP genotyping with the 777K SNP chip showed that 9.1% of loci on the chip are polymorphic in the herd, compared with 62–90% seen in commercial cattle breeds. Instead of being homogeneously distributed along the genome, these loci are clustered at specific chromosomal locations. A high proportion of the Chillingham individuals examined were heterozygous at many of these polymorphic loci, suggesting that some loci are under balancing selection. Some of these frequently heterozygous loci have been implicated as sites of recessive lethal mutations in cattle. Linkage disequilibrium equal or close to 100% was found to span up to 1350 kb, and LD was above r² = 0.25 up to more than 5000 kb. This strong LD is consistent with the lack of polymorphic loci in the herd. The heterozygous regions in the Chillingham cattle may be the locations of genes relevant to fitness or survival, which may help elucidate the biology of local adaptation in traditional breeds and facilitate selection for such traits in commercial cattle.     

Genetics of Lipid-Storage Management in Caenorhabditis elegans Embryos

Lipids play a pivotal role in embryogenesis as structural components of cellular membranes, as a source of energy, and as signaling molecules. On the basis of a collection of temperature-sensitive embryonic lethal mutants, a systematic database search, and a subsequent microscopic analysis of >300 interference RNA (RNAi)–treated/mutant worms, we identified a couple of evolutionary conserved genes associated with lipid storage in Caenorhabditis elegans embryos. The genes include cpl-1 (cathepsin L–like cysteine protease), ccz-1 (guanine nucleotide exchange factor subunit), and asm-3 (acid sphingomyelinase), which is closely related to the human Niemann-Pick disease–causing gene SMPD1. The respective mutant embryos accumulate enlarged droplets of neutral lipids (cpl-1) and yolk-containing lipid droplets (ccz-1) or have larger genuine lipid droplets (asm-3). The asm-3 mutant embryos additionally showed an enhanced resistance against C band ultraviolet (UV-C) light. Herein we propose that cpl-1, ccz-1, and asm-3 are genes required for the processing of lipid-containing droplets in C. elegans embryos. Owing to the high levels of conservation, the identified genes are also useful in studies of embryonic lipid storage in other organisms.