Transformation of plant young proembryos by electroporation

It is first reported that plant young proembryos expressed exogenous reporter genes by electroporation. Young proembryos with 8–32 cells and globular proembryos with 250–400 cells could be isolated by enzymatic maceration combined with microdissection. After electroporation withGUS orGFP genes, the proembryos were cultured for 1–2 d in KM8p medium. At the field strength of electroporation 500–1500 V/cm, blue reaction of GUS or green fluorescence of GFP could be observed in the proembryos. The highest transient expression frequency of young proembryos (2.2%) was obtained at the field strength of 750 V/cm, whereas the highest frequency of globular proembryos (5.9%) was obtained at the field strength of 1 250 V/cm. Taking the proportion of transformed cells in the whole cells of proembryos as efficient transformation frequency, the efficient transformation frequency of the young proembryos was 7 times that of the globular proembryos.     

板栗空篷的蛋白质和氨基酸含量变化的研究

本文研究了空篷栗、正常栗的总苞和子房、子叶发育过程中蛋白质和氨基酸含量的变化。结果表明：空篷栗在幼胚发育期、子叶发育期，其总苞和子房中的蛋白质含量均低于正常栗；空篷栗在传粉受精后，子房中的氨基酸含量均低于正常栗；子房、子叶中的１８种氨基酸含量均高于总苞。总苞发育过程中，丙氨酸和异亮氨酸的含量空篷栗低于正常栗；子房、子叶发育过程中，色氨酸、异亮氨酸、苏氨酸、甘氨酸、丝氨酸和赖氨酸的含量空篷栗低于正常栗。这表明，在幼胚发育、子叶发育期，空篷栗得不到足够的物质供应，影响物质的定向运输和积累，导致了空篷栗的产生。文中还讨论了谷氨酸、天门冬氨酸、色氨酸、脯氨酸的含量变化与子房、子叶发育的关系。     

莼菜绒毡层细胞发育的超微结构变化

莼菜（Ｂｒａｓｅｎｉａ ｓｃｈｒｅｂｅｒｉＧｍ ｅｌ）绒毡层细胞在小孢子母细胞时期,质体出现变形,淀粉粒消失．减数分裂期,绒毡层细胞中有丰富的高尔基体,内质网,核糖体和线粒体．随着细胞的发育,细胞壁呈胶质状态,胞间连丝断离,细胞间发育出现不同步现象,核糖体开始解体形成电子密度高的球状体．单核小孢子时期,质体积累淀粉增多,细胞内部微结构开始解体．经观察,绒毡层细胞壁的融解物质参与了小孢子外壁的形成,核糖体形成的高电子密度球状体沉积小孢子外壁腔隙中形成外壁蛋白．绒毡层细胞中淀粉粒积累与消失是小孢子正常发育的必然结果．     

莲子对黄曲霉毒素（B_1）诱发的人体淋巴细胞姐妹染色单体互换和细胞周期状态的影响

莲子对黄曲霉毒素（Ｂ_１）诱发的人体淋巴细胞姐妹染色单体互换和细胞周期状态的影响干侣仙，廖绵初，黄少珍（厦门大学抗癌研究中心，广西壮族自治区肿瘤研究所）肿瘤细胞遗传学和肿瘤形成过程的研究已表明［１］在细胞恶性转化和肿瘤形成的多阶段过程中，存在着突变，...     

华南五针松幼茎发育研究

利用透射电子显微镜和光学显微镜对华南五针松(Pinuskwangtungensis)顶芽和一年生幼茎,进行形态结构和发育的观察,并进行细胞内淀粉粒含量的研究。结果表明,华南五针松幼茎在芽鳞形成后期,顶芽出现分区结构。顶芽细胞、幼茎细胞中,质体、线粒体、内质网等细胞器的分布、数量与其干旱生活的环境相适应。茎的皮层、初生木质部有树脂道分布,均为裂生型垂直树脂道,由莲座状排列的12～18个细胞发育而来。淀粉粒在树脂道发育成熟前后的动态变化与树脂的合成和分泌有一定的关系。随着各组织的发育完善,幼茎内薄壁细胞的淀粉粒呈现一个由无到丰富的动态变化。     

莲子叶蛋白体的类型,来源与发育

莲子叶于发育的１０￣２８ｄ,以不同速率在叶肉细胞中大量积累贮存蛋白,并以不同方式组装许多形态各异、结构不同的蛋白体。按其来源与发育的特点可分４种类型：①粗面球状蛋白体;②光面球状蛋白体;③杯状蛋白体;④淀粉蛋白体。发育顺序是①,②类先形成,③,④类后发生;②类的发育贯穿整个贮存蛋白的积累期,比较了莲与其他植物蛋白体的异同。     

Preliminary study of microscale zircon oxygen isotopes for Dabie-Sulu metamorphic rocks： Ion probe in situ analyses

151 in situ analyses of oxygen isotopes were carried out by ion micro-probe for zircons from 8 localities of HP-UHP metamorphic rocks including eclogites in the Dabie-Sulu terrane. The results show significant heterogeneity in δ^18O values, with variation in different rocks from -8.5‰ to 9.7‰ and within one sample from 2‰ to 12‰. No measurable difference in δ^18O was observed between protolith magmatic (detrital) zircons and metamorphic recrystallized zircons within analytical uncertainties from the ion micro-probe measurements. This indicates that the metamorphic zircons have inherited the oxygen isotopic compositions of protolith zircons despite the HP to UHP metamorphism. According to their protolith ages from zircon U-Pb in situ dating by the same ion micro-probe, two groups of oxygen isotope composition are recognized, with one having δ^18O values of 6‰-7‰ for old protolith of 1.9-2.5 Ga ages and the other 0‰-2‰ for young protolith of 0.7-0.8 Ga ages. The latter anomalously low δ^18O values of zircons indicate that the magma has had the obvious involvement of meteoric water when forming the young protolith of high-grade metamorphic rocks. This may be correlated with the snowball Earth event occurring in South China and the world elsewhere during the Neoproterozoic.     

Rice protoplasts isolated directly from mature-embryo-derived calli

Mature-embryo-derived calli of japonica rice (Oryza sativa L) Taipei 309 were used for replicated protoplast isolation experiments. Six out of nine callus lines produced protoplasts with satisfactory yield of 5.20×10⁶–8.96×10⁶ protoplasts/g FW (fresh weight). The remaining three callus lines initiated from seeds of cryopreserved-callus-derived plants had rooty calli, resulting in low yield of protoplasts and a large number of isolated banana-shape intact cells. Viability of protoplasts ranged 87.46%–94.15%. The average size of protoplasts was 207.49–379.04 μm² in different callus lines. Comparitive experiments were also carried out using both calli and suspension culture cells for protoplast isolation. The results demonstrated that protoplast isolation of calli was a substantially simplified and reliable method for preparing rice protoplasts. Jin Deming: born in Jan. 1959, Associated Professor     

Regenerating plants from cryopreserved adventitious buds of haploids in rice

A large number of adventitious buds were induced fromin vitro cultured young inflorescences of haploids of rice. Having been subcultured on solidified subculture media at 26°C for 7 days, the adventitious buds were loaded into 1.8 mL plastic cryotubes with cryoprotectant and kept on ice for 45–60 min. After cooled at a rate of 1.0°C/min down to −40°C, the samples, were kept in liquid nitrogen. The adventitious buds which have been cryopreserved for about 30 days were thawed rapidly in 38–40°C water and then plated on solidified MS medium containing 3% sucrose, 0.5 mg/L NAA and 2.0 mg/L kinetin. After plated, 23%–32% of adventitious buds resumed growth and 15%–22% regenerated plantlets. The results of this work indicated that the adventitious buds derived fromin vitro cultured young inflorescences is a critical factor for the success and subculturing adventitious buds on MS medium containing 3% sucrose and 4% sorbitol or 20% potato extract is essential to the procedure. The effective cryoprotectant is 10% DMSO (dimethyl sulfoximide)+0.5 mol/L sorbitol. Supported by the Natural Science Foundation of Hubei Province Zhang Zhihong: born in 1963, Lecturer     

发根农杆菌Ri质粒诱导的黄芪发状根根冠细胞亚显微结构研究

对膜荚黄芪发状根根冠细胞的亚显微结构进行研究发现：发状根根尖没有分化出典型的根冠结构；根冠平衡囊细胞中淀粉分布无极性，散地细胞质中；细胞中未发现粗面内质网而仅有滑面内质网的存在；高尔基体分泌功能不旺盛，无肥大的潴泡；细胞壁较薄，纵向壁略加厚，其间无粘液积累；根冠外周细胞逐渐老化，但细胞壁不溶解，老化的细胞不脱落，且根冠细胞层数不增加。     

Effect of monoclonal antibody to bull sperm sp18 on murine fertilization and embryos development

Western blot analysis revealed that one IgG1 monoclonal antibody (mAb) to sp18 family membrane proteins (Mr. 14, 16 and 18 ku) of bovine sperm reacted faintly with protein bands of 14, 18, 22, 30 and 60 ku (reducing) in samples of mouse sperm. The mAb also reacted to protein of egg lysozyme. Using a laser confocal microscope, indirect immunofluorescence (IIP) showed that the sp18 antigens were present in the posterior head of murine sperm. In murine in vitro fertilization (IVF) and embryo development trails, a total of 426 oocytes from C57BL/6 and F1 hybrid strain (CD1 × C57BL/6 cross) of 12 female mice were used in 3 independent trails. After preincubating capacitated sperm with 182 μg/mL of sp18 mAb in the modified TYH IVF medium for 15-20 min, cumulus-oocyte-complexes were introduced. The fertilization rate in sp18 mAb groups was 77.1 %, which was not significantly (P > 0.05) different from the nonspecific mouse IgG (79.2%) and non-IgG (80.3 %) control groups. Fertilized oocytes had been continuously cultured in modifed CZB medium. 100%, 100% and 97.9% of 1-cell embryos developed to 2-cell stage in sp18 mAb, nonspecific mouse IgG and non-IgG group 30 h after the start of fertilization, respectively. In the nonspecific mouse IgG and non-IgG groups, 64.1 % and 64.3% of embryos developed to the 4-cell stage, respectively, but all developing eggs in sp18 mAb groups arrested development in vitro at 2-cell stage. After zonae of 2-cell blocked embryos were enzy-matically removed with 0.5% pronase, detection of sp18 antigens by IIF indicated that the fluorescence scattered on two embryonic cells. For embryos fertilized in vivo and co-cultured with 182 μg/mL sp18 mAb, the numbers of 1-cell embryos reaching the 2-cell and 4-cell stage were 95. 2% and 70. 5%, which were not significantly (P>0.05) different from the control group (92.9% and 77.9%). These results indicate that the sp18 antigens on posterior head of mouse sperm were incorporated into the egg plasma membrane during fertilization, and played an active role in development of murine preimplanta-tion embryo.     

Effect of internal deletion of Myosin II on growth and development ofDictyostelium discoideum

Four deletion mutantDictyostelium myosin II heavy chain genes, MyΔ824-941 (Δ1/ 3S2), MyΔ934-1454 (ΔS2), MyΔ934-1194 (ΔS2-1) and MyΔ1 157–1454 (ΔS2-2), were transformed by standard electfoporation into mhcA-cells (T-null), a mutantDictyostelium cell devoid of endogenous myosin II heavy chain gene. The growth, development and formation of fruiting bodies of cells expressing those mutant myosin II s under suspension culture were investigated by comparison with the wild type cell. The results indicate that internal deletion of myosin II affeds the growth and development ofDictyastelium. Furthermore, the longer the length of deletion, the more serious the defect in phenotype.     

Electrophysiological research of synapse formation from beginning to maturity

The development of synapses is an important question in neurobiology. We employ patchclamp technique to record MEPCs and EPCs of myoballs inXenopus cell culture. The rate of EPC/MEPC is related with synapses maturation. Accordingly, we define the mature synapses as those whose rate of presynaptic membrane ’ s EPC to MEPC is more than 8. The process of maturation needs about 2–5 h after neuntes contact with myoballs. We divide the development of synapses into three stages: young, developing and mature. The neurotransmitters stack gradually and increase by hundreds of times along with young synapses growing into maturity. So the process of synapses maturation should include presynaptic membrane specialization, postsynaptic membrane specialization and neurotransmitters stack.     

莲子贮存蛋白基因的表达与子叶叶肉细胞的变构

莲子贮存蛋白基因（ＳＰＧ）的表达受发育调控。子叶叶肉细胞变构早于ＳＰＧ的开始表达,随发育过程加剧。在贮存组织中细胞变构的发生有严格的时空顺序性。变构主要表现在细胞核体积增大,外形变不规则,染色质分布不均匀化,核膜局部消失。     

QTL analysis of rice low temperature germinability

A double haploid population, derived from anther culture of F1 hybrid between a typical indica and a japonica (ZYQ8/JX17), has been used to investigate the low temperature germinability (LTG) at 15 ℃. The low temperature germinability of two parents was significantly different. In 6-11 d, the germination percentage of ZYQ8 was higher than that of JX17. In 12-16 d, the germination percentage of JX17 was higher than that of ZYQ8. The quantitative trait loci (QTLs) of every day for low temperature germinabilityhave been mapped based on a molecular linkage map constructed from this population. In 8-11 d, qLTG-9 was identified in C397B-RZ617B on chromosome 9, the additive effect was positive, showing that, the allele from JX17 could increase low temperature germinability. In 12-16 d, qLTG-4 was mapped between RG908 and CT563 on chromosome 4, the additive effect was negative, showing that the allele from ZYQ8 could increase low temperature germinability. These two QTLs were detected at different stages, showing the complexity of the mechanism of low temperature germinability.     

Cloning and characterization of a gene encoding cysteine proteases from senescent leaves of Gossypium hirsutum

A gene encoding a cysteine proteinase was isolated from senescent leave of cotton (Gossypium hirsutum) cv liaomian No. 9 by utilizing rapid amplification of cDNA end spolymerase chain reaction (RACE-PCR), and a set of consensus oligonucleotide primers was designed to anneal the conserved sequences of plant cysteine protease genes. The cDNA, which designated Ghcysp gene, contained 1368 bp terminating in a poly(A)^ trail, and included a putative 5‘(98 bp) and a 3‘(235 bp) non-coding region. The opening reading frame (ORF) encodes polypeptide 344 amino acids with the predicted molecular mass of 37.88 kD and theoretical pl of 4.80. A comparison of the deduced amino acid sequence with the sequence in the GenBank database has shown considerable sequence similarity to a novel family of plant cysteine proteases. This putative cotton Ghcysp protein shows from 67% to 82% identity to the other plants. All of them share catalytic triad of residues, which are highly conserved in three regions. Hydropaths analysis of the amino acid sequence shows that the Ghcysp is a potential membrane protein and localizes to the vacuole, which has a transmembrane helix between resides 7-25. A characteristic feature of Ghcysp is the presence of a putative vacuole-targeting signal peptide of 19-amino acid residues at the N-terminal region. The expression of Ghcysp gene was determined using northern blot analysis. The Ghcysp mRNA levels are high in development senescent leaf but below the limit of detection in senescent root, hypocotyl, faded flower, 6 d post anthesis ovule, and young leaf.     

Molecular cloning of a fulllength cDNA for ECBP21 from Angelica dahurica

ECBP21 is an extracellular calmodulin-binding protein which was first detected and purified from extracellular extracts of suspension-cultured cells of Angelica dahurica. The purified protein was electroblotted onto PVDF membrane and the amino acid sequences from 1 to 20 were determined. Using degenerate oligonucleotides of the sequence, a full-length cDNA coding for ECBP21 was isolated by a combination of RT-PCR and 5′-RACE cloning. The cDNA contains 947 nucleotides and codes for a precursor protein of 216 amino acids. The N-terminal 1–25 amino acid sequence is a predicted signal peptide and the other 26–216 amino acid sequence is a mature peptide. The 26–45 amino acid sequence shows identity with the N-terminal amino acid sequence of purified ECBP21 from Angelica dahurica. The fragment of encoding the mature protein was cloned into pET-28b(+) and transformed into E. coli BL21(DE3). A protein with relative molecular mass 21 ku was expressed in E. coli. Using a biotinylated-CaM gel overlay technique, the expression protein was tested for its ability to bind CaM. The results indicated that the expression protein is a Ca²⁺-dependent CaM-binding protein. Thus, these results further defined the cDNA clone for ECBP21. This work laid a foundation for elucidating biological functions of ECBP21 by using molecular biological means.     

Genome organization and phylogenetic tree analysis of Garlic virus E, a new member of genus Allexivirus

The complete sequence of an Allexivirus isolated from garlic plants in Yuhang City, Zhejiang Province, China had been determined. The single-strand, positive RNA genome was 8451 nucleotides in length excluding poly(A) tail. The genome organization of this virus was similar to that of the other Allexiviruses but only with 62.8%–64.8% nucleotide acid identities. The amino acid sequences of proteins encoded by ORF1-6 shared 67.6%–78.5%, 55.4%–66.2%, 56.7%–66.4%,40.3%–55.6%,66.3%–79.7%and 52.2%–68.8% identities with those of the others respectively. The homology range between it and the other Allexiviruses was similar to that between the other distinct species in this genus. A more comprehensive comparison using all available CP amino acid sequences showed that it shared only 63.9%–79.8% amino acids identical with the others. Therefore, it had been considered as a new member of the genus, named as garlic virus E (GarV-E). Phylogenetic analysis confirmed GarV-E as a distinct member and the correct names and classification of some members of genus Allexivirus were also discussed.     

Comparison of effective parameters for copper powder production <Emphasis Type="Italic">via</Emphasis> electrorefining and electrowinning cells and improvement using DOE methods

The influences of cupric ion concentration (5–35 g/L), current density (500–2000 A/m²), circulation rate of the electrolyte solution (15–120 mL/min), and temperature (25–60°C) on the physical and chemical properties of copper powders obtained in electrolysis cells were investigated. Two industrial processes, electrorefining (ER) cells with a synthetic electrolyte and electrowinning (EW) cells with an original solution of copper mineral leaching, were utilized to produce copper powders. Finally, the statistical full factorial method of design of experiments (DOE) was employed to investigate the interaction or the main effects of processes. The results show that increasing the copper concentration and temperature can increase the grain size, apparent density, and electrical energy consumption. On the other hand, increasing the current density and circulation rate of the electrolyte can decrease them. This production process is optimized via DOE to control the interactive and main effects to produce copper powders with favorable properties.     

甘薯小孢子发生的超微结构观察

甘薯小孢子发生过程中，细胞质超微结构发生了显著的变化，其变化主要涉及小泡，内质网质和核糖体，小泡在减数分裂过程中尤为丰富，参与胼胝质壁的形成，内质网在小孢子母细胞中非常丰富，在减数分裂过程中转化为小泡，在小孢子是时期转化为内质网小泡，质体在小孢子母细胞中比较丰富，但处于退化状态，到小孢子时期尤为丰富并集中分布在细胞核的周围，核糖体的密度在小孢子母细胞和小孢子中均较富，在减数分裂过程中降低。