Cloning, Characterization, and Expression Analysis of Calreticulin from Pearl Oyster Pinctada fucata

Calreticulin is a unique calcium-binding protein with multiple functions mostly located in the sarcoplasmic/endoplasmic reticulum. A large amount of calcium is absorbed from the medium and transported to mineralization sites during biomineralization in pearl oyster. This paper describes the cloning of the full-length cDNA of calreticulin from Pinctada fucata, namely PCRT. PCRT encodes a deduced 414-amino acid protein, which includes a predicted 17- amino acid signal peptide and an endoplasmic reticulum retrieval sequence HDEL. The protein shows 63%-76% sequence identity and shares some common characteristics with calreticulins from other species. Semi-quantitative RT-PCR indicates that PCRT is ubiquitously expressed in all tissues tested with the highest expression in the hemolymph and the mantle. In situ hybridization analysis of PCRT in the mantle showed strong signals in the inner fold, the inner side of middle fold, and the inner side of outer fold of the mantle epithelium, All these results suggest PCRT might be involved in Ca^2＋ transport and storage during oyster biomineralization.     

Cloning, Characterization, and Expression Analysis of Calreticulin from Pearl Oyster Pinctada fucata

Calreticulin is a unique calcium-binding protein with multiple functions mostly located in the sar-coplasmic/endoplasmic reticulum.A large amount of calcium is absorbed from the medium and transported to mineralization sites during biomineralization in pead oyster.This paper describes the cloning of the full-length cDNA of calreticulin from Pinctada fucata,namely PCRT.PCRT encodes a deduced 414-amino acid protein,which includes a predicted 17- amino acid signal peptide and an endoplasmic reticulum retrieval sequence HDEL.The protein shows 63%-76% sequence identity and shares some common characteristics with calreticulins from other species.Semi-quantitative RT-PCR indicates that PCRT is ubiquitously ex-pressed in all tissues tested with the highest expression in the hemolymph and the mantle.In situ hybridiza-tion analysis of PCRT in the mantle showed strong signals in the inner fold,the inner side of middle fold,and the inner side of outer fold of the mantle epithelium.All these results suggest PCRT might be involved in Ca2+ transport and storage during oyster biomineralization.     

Localization of calmodulin and calmodulin-like protein and their functions in biomineralization in P. fucata

Calmodulin (CaM) and calmodulin-like protein (CaLP) are two proteins involved in biomineralization. Their localizations in Pinctada fucata mantle epithelia were studied by Western blot (WB) analysis of the nuclear/cytosol fraction of primary cultured Pinctada fucata mantle cells and immunogold electron microscopy. The results showed a completely different distribution of these two proteins at the subcellular level. CaM was distributed throughout both the nucleus and cytoplasm of the mantle epithelium but CaLP was distributed only in the cytoplasm. The functions of these two proteins in biomineralization were investigated by shell regeneration. During this process, the expressions of CaM and CaLP were greatly enhanced in different organelles of the mantle epithelium. Overexpression of these two proteins and a mutant of calmodulin-like protein (M-CaLP) that lacks an extra C-terminal tail in MC3T3-E1 promoted the mRNA expression of osteopontin, a biomineralization marker for osteoblasts. All of the results indicated that CaM and CaLP have completely different distributions in the mantle epithelium and affect the biomineralization process at different levels. The extra C-terminal tail of CaLP is important for its functions in biomineralization in Pinctada fucata.     

Cloning and characterization of 37 kD laminin receptor precursor in pearl oyster, Pinctada fucata

A 1063bp cDNA clone encoding a putative 37 kD laminin receptor precursor (37 kD LRP) is isolated from the mantle tissue of pearl oyster, Pinctada fucata. The amino acid sequence predicted from the cDNA sequence is 301 residues long, with a calculated molecular mass of 33.5 kD. RT-PCR analysis shows that 37 kD LRP mRNA is especially highly expressed in the mantle while widely expressed in several tissues. In situ hybridization analysis reveales that 37 kD LRP is expressed in the outer epithelial cells of the mantle edge, suggesting its involvement in cell proliferation and secretion in P. fucata. The identification and characterization of 37 kD LRP in the pearl oyster will help us to further understand the signal transduction in the processes of mantle epithelial cell proliferation and tissue formation.     

锯缘青蟹不同器官组织中4种类型ATPase活性比较研究

本实验采用生化方法研究了锯缘青蟹的鳃、肝胰腺和肌肉等不同器官组织中4种类型ATPase活性.结果表明,不同器官组织中同类型ATPase活性各异,Na K ATPase活性由强至弱为鳃>肝胰腺>肌肉.Ca2 ATPase活性为肌肉>肝胰腺>鳃.Mg2 ATPase活性为鳃>肌肉>肝胰腺.Ca2 Mg2 ATPase活性则为肌肉>鳃>肝胰腺.同一器官组织中不同类型ATPase活性也有较大的差异,鳃中Na K ATPase活性最大,其余的依次为Mg2 AT Pase,Ca2 Mg2 ATPase,Ca2 ATPase;肝胰腺中Na K ATPase活性最强,其余的依次为Ca2 Mg2 ATPase,Mg2 ATPase,Ca2 ATPase;肌肉中Ca2 Mg2 ATPase活性最大,其余的依次为Ca2 ATPase,Na K ATPase,Mg2 ATPase.此外,锯缘青蟹同一器官组织中同类型ATPase在不同个体间也存在一定差异.不同器官组织中4种类型ATPase活性各异主要与其所执行的生理功能密切相关.     

Morphological and physiological analysis of narrow and striped leaf 1 （nsl1） mutant of rice （Oryza sativa L.） and the gene mapping

The shape and color of rice leaves are impor- tant agronomic traits that directly influence the proportion of sunlight energy utilization and ultimately affect the yield and quality. A new mutant exhibiting stable inheritance was identified as derived from ethyl methane sulfonate （EMS）-treated restorer Jinhui 10, tentatively named as narrow and striped leaf 1 （nsll）. The nsll displayed pale white leaves at the seeding stage and then white striped leaves in parallel to the main vein at the jointing stage. Meanwhile, its leaf blades are significantly narrower than the control group of Jinhui 10. The chloroplast structures of cells in the white striped area of the nsll mutant break down, and the photosynthetic pigments are significantly lower than that of the wild type. Moreover, fluorescence parameters, such as Fo, Fv/Fm, ФpsⅡ, qP, and ETR, in the nsll mutant are significantly lower than those of the wild type, and the photosynthetic efficiency is also significantly decreased. These changes in leaf color and shape, together with physiological changes in the nsll, result in smaller plant height and a decrease in the most important agro- nomic traits, such as the number of grains per panicle, grain weight, etc. Genetic analysis shows that the narrow and striped traits of the nsll mutant are controlled by a single recessive nuclear gene, which is located between InDel 16 and InDel 12 in chromosome 3. The physical distance is 204 kb. So far, no similar genes of such leaf color and shape in this area have been reported, This study has laid asolid foundation for the gene cloning and function analysis of NSL 1.     

Genetic analysis and fine mapping of the pubescence gene GL6 in rice (Oryza sativa L.)

The pubescence of the leaf blade surface is an important agronomic characteristic for rice morphology and significantly influences rice growth as well as physiological characteristics. This characteristic was analyzed in F1 and F2 plants derived by crossing cultivar 75-1-127 with the indica cultivar Minghui 63, as well as the glabrous cultivar Lemont and indica cultivar 9311. Results indicated that the pubescence of the leaf blade surface was a dominant trait and controlled by a single gene. The GL6 gene was primarily mapped on rice chromosome 6 with recessive F2 population derived from 75-1-127/Minghui 63 by combining bulked segregation analysis and recessive class analysis using the Mapmaker3.0/MapDraw software. The genetic distances between the simple sequence repeat markers RM20491 and RM20547 were 7.2 and 2.2 cM, respectively. The GL6 gene was fine mapped in the interval between InDel-106 and InDel-115 at genetic distances of 0.3 and 0.1 cM, respectively. The large, recessive F2 population was derived from 75-1-127/Minghui 63. A high-resolution genetic and physical map of GL6 was constructed. Derived from the map-based sequences published by the International Rice Genome Sequencing Project, the GL6 gene was localized at an interval of 79 (japonica) and 116.82 kb (9311) bracketed by InDel-106 and InDel-115 within the BAC accession numbers AP008403 and AP005760. Seven annotated genes (japonica) and eight annotated genes (9311) were present. The basis was further set for GL6 cloning and function analysis.     

Thermoluminescence of Al2O3 crystals grown by temperature gradient techniques

Color center concentrations of the Al2O3 crystal grown by temperature gradient techniques were calculated from Gaussian fits to absorption spectra. The concentrations for F- and F + -centers at 204, 232, and 255 nm were determined to be 1.361×10 17 , 0.098×10 17 , and 0.325×10 17cm-3 , respectively. Studies have shown that the thermoluminescence (TL) glow curve exhibits a prominent 450 K peak that matches well the first-order fitting curve. The thermal activation energy E and frequency factor s of the trap were determined to be 0.94456±0.00545 eV and 5.8703×10 11 s-1 . With this theoretical analysis, a simple one-trap/one-center TL model is presented to provide a theoretical explanation of the TL process.     

Construction of a linkage map and QTL mapping for fiber quality traits in upland cotton (Gossypium hirsutum L.)

With the development in spinning technology, the improvement of cotton fiber quality is becoming more and more important. The main objective of this research was to construct a high-density genetic linkage map to facilitate marker assisted selection for fiber quality traits in upland cotton (Gossypium hirsutum L.). A genetic linkage map comprising 421 loci and covering 3814.3 cM, accounting for approximately 73.35% of the cotton genome, was constructed using an F2 population derived from cross GX1135 (P 1 )×GX100-2 (P 2 ). Forty-four of 49 linkage groups were assigned to the 26 chromosomes. Fiber quality traits were investigated in F2 population sampled from individuals, and in F2:3 , and F2:4 generations sampled by lines from two sites and one respectively, and each followed a randomized complete block design with two replications. Thirty-nine quantitative trait loci were detected for five fiber quality traits with data from single environments (separate analysis each): 12 for fiber length, five for fiber uniformity, nine for fiber strength, seven for fiber elongation, and six for fiber micronaire, whereas 15 QTLs were found in combined analysis (data from means of different environments in F2:3 generation). Among these QTLs, qFL-chr5-2 and qFL-chr14-2 for fiber length were detected simultaneously in three generations (four environments) and verified further by combined analysis, and these QTLs should be useful for marker assisted selection to improve fiber quality in upland cotton.     

Localization of calmodulin and calmodulin-like protein and their functions in biomineralization in P. fucata

Calmodulin （CAM） and calmodulin-like protein （CaLP） are two proteins involved in biomineralization. Their localizations in Pinctada fucata mantle epithelia were studied by Western blot （WB） analysis of the nuclear/cytosol fraction of primary cultured P. fucata mantle cells and immunogold electron microscopy. The results showed a completely different distribution of these two proteins at the subcellular level. CaM was distributed throughout both the nucleus and cytoplasm of the mantle epithelium but CaLP was distributed only in the cytoplasm. The functions of these two proteins in biomineralization were investigated by shell regeneration. During this pro- cess, the expressions of CaM and CaLP were greatly enhanced in different organelles of the mantle epithelium. Overexpression of these two proteins and a mutant of calmodulin-like protein （M-CaLP） that lacks an extra C-terminal tail in MC3T3-E1 promoted the mRNA expression of osteopontin, a biomineralization marker for osteoblasts. All of the results indicated that CaM and CaLP have completely different distributions in the mantle epithelium and affect the biomineralization process at different levels. The extra C-terminal tail of CaLP is important for its functions in biomineralization in P. fucata.     

Research on a Class of Equations over Finite Fields

Let F_q stand for the finite field of odd characteristic p with q elements(q=p~n,n∈N)and F_q~* denote the set of all the nonzero elements of F_q.In this paper,by using the augmented degree matrix and the result given by Cao,we obtain a formula for the number of rational points of the following equation over F_q:f(x _1,x _2,...,x _n)=(a_1 x_1 x_2~d+a_2 x_2 x_3~d...+a_(n-1)x_(n-1)x_n~d+a_n x_n x_1~d)~λ-bx_1~(d1)x_2~d2...x_n~(dn),with a_i,b∈F_q~*,n≥2,λ0 being positive integers,and d,d_i being nonnegative integers for 1≤i n.This technique can be applied to the polynomials of the form h_1~λ=h_2 with λ being positive integer and h_1,h_2∈F_q[x _1,x _2,...,x _n].It extends the results of the Markoff-Hurwitz-type equations.     

Zircon U/Pb dating and Hf-O isotopes of the Zhouan ultramafic intrusion in the northern margin of the Yangtze Block, SW China: Constraints on the nature of mantle source and timing of the supercontinent Rodinia breakup

The Zhouan ultramafic intrusion in the northern margin of the Yangtze Block is mainly composed of lherzolite.Zircon grains selected from lherzolite are irregular in shape with distinct oscillatory and sector zoning and have Th/U ratios ranging from 0.8 to 10.6,indicating a magmatic origin.The weighted average 206 Pb/238 U age is 637±4 Ma (2σ,n=15),which can be considered as the crystallization age of the Zhouan intrusion.Zircon grains have δ18O values ranging from 5.2‰ to 7.0‰,with an averaged value of 5.8±0.4‰ (1σ,n=33),similar to the mantle δ18O value of zircon.Their 176Hf/177Hf(t) ratios range from 0.282410 to 0.282594 with εHf(t) values ranging from 1.3 to 7.6,lower than the corresponding value of the depleted mantle (～15),indicating an enriched mantle source.The enriched mantle source may have generated from a metasomatized lithospheric mantle with subducted slab.A number of ～635 Ma mafic-ultramafic intrusions in the Suizao basin are associated with coeval bimodal volcanics of the Yaolinghe Formation,indicating a continental rift setting.The ～635 Ma magmatic event in this region may represent the product of the last breakup of the Rodinia supercontinent in the northern margin of the Yangtze Block at Neoproterozoic.     

Characteristics of lateral heterogeneities with thermal and chemical origins in the pyrolitic lower mantle

The relative changes between shear and compressional velocities (R_SP = ∂ ln V_S/∂ ln V_P), bulk sound and shear velocities (R_CS = ∂ ln V_C/∂ ln V_S), and density versus shear wave velocity (R_ρS = ∂ ln ρ/∂ ln V_S) in response to thermal and chemical variations were investigated for the pyrolitic lower mantle. For heterogeneities with thermal origins, R_SP increases from 1.7 to 2.0 together with R_ρS decreasing from 0.4 to 0.2 and R_CS = ∼0.27 from the top to the bottom of the lower mantle. In comparison, chemical variations (bulk iron or silica contents) are characterized by R_SP < 1.5 and R_CS > 0.5 at lower mantle depths. Negative values of R_ρS and R_CS are indicative of chemical anomalies in the lower mantle, but a combination of thermal and chemical heterogeneities may be required to produce velocity and density anomalies at the magnitudes observed in seismic data. Further refinement of these characteristics requires data on the higher order pressure and temperature derivatives of the elastic moduli of the constituent phases.     

Change of proton motive force across thylakoid membrane in soybean leaf during state transitions

Change of proton gradient across thylakoid membrane in soybean leaves was studied with millisecond delayed light emission(ms-DLE) during the coures of state transitions which were indicated by the cholrophyll fluores-cence at room temperature and 77K.When dark-adapted leaves were induced to state Ⅰ with far-red light,Fm/F0,F685/F735 and the intensity of fast phase of ms-DLE were af-fected slightly.However,during the induction to state Ⅱ with red light,both Fm/F0 and F685/F735 decreased immedi-ately and the former were quicker than the latter.In this interval,the intesity of fast phase of ms-DLE increased to a maximum and then decreased to a lower valus during the transition to stateⅡ.Nigericin,an uncoupler which elimi-nates the proton gradient across thylakoid membrane,inhib-ited the increase in the intensity of fast phase of ms-DLE during the transition to state Ⅱ.Another uncoupler,valino-mycin,which eliminates the membrane potential,did not affect the changes of the intensity of fast phase.These results suggest that the prompt increase in the intensity of fast phase of ms-DLE at the beginning of transitions to state Ⅱ is cor-related mainly with the proton gradient released from water oxidation in photosystemⅡ.     

Cloning and characterization of 37 kD laminin receptor precursor in pearl oyster, Pinctada fucata

A 1063 bp cDNA clone encoding a putative 37 kDa laminin receptor precursor （37 kDa LRP） is isolated from the mantle tissue of pearl oyster, Pinctadafucata. The amino acid sequence predicted from the cDNA sequence is 301 residues long, with a calculated molecular mass of 33.5 kDa. RT-PCR analysis shows that 37 kDa LRP mRNA is especially highly expressed in the mantle while widely expressed in several tissues. In situ hybridization analysis reveals that 37 kDa LRP is expressed in the outer epithelial ceils of the mantle edge, suggesting its involvement in cell proliferation and secretion in P. fucata. The identification and characterization of 37 kDa LRP in the pearl oyster will help us to further understand the signal transduction in the processes of mantle epithelial cell proliferation and tissue formation.     

Cloning and characterization of 37 kDa laminin receptor precursor in pearl oyster, Pinctada fucata

A 1063 bp cDNA clone encoding a putative 37 kDa laminin receptor precursor (37 kDa LRP) is isolated from the mantle tissue of pearl oyster, Pinctadafucata. The amino acid sequence predicted from the cDNA sequence is 301 residues long, with a calculated molecular mass of 33.5 kDa. RT-PCR analysis shows that 37 kDa LRP mRNA is especially highly expressed in the mantle while widely expressed in several tissues. In situ hybridization analysis reveals that 37 kDa LRP is expressed in the outer epithelial cells of the mantle edge, suggesting its involvement in cell proliferation and secretion in P. Fucata. The identification and characterization of 37 kDa LRP in the pearl oyster will help us to further understand the signal transduction in the processes of mantle epithelial cell proliferation and tissue formation.     

LiNiPO4前驱体固相反应合成动力学及电容性能

以氢氧化锂、磷酸二氢铵和醋酸镍为原料,以聚乙二醇PEG-400为表面活性剂,采用前驱体固相活化法制备LiNiPO4纳米晶材料.固相制备过程的活化能E=81.08 kJ·mol-1,过程动力学为二维相界面扩散反应机理.前驱体生成LiNiPO4的反应符合界面反应指数成核机理,活化能E1=11.82 kJ·mol-1,指前因子lnA=13.82;LiNiPO4晶体生长过程具有较小的活化能E2=17.73 kJ·mol-1;Li3PO4转化反应和反应体系物系晶化过程符合二维相界面扩散反应机理,其是制备过程可控制的重要步骤.材料复合电极LiNiPO4+Nafion/C在0.5 mol·L-1H2SO4中具有典型的电容性能,电极比电容为214 F·g-1,经1 000次循环,电极电容量不但没有衰减反而略有增加,是潜在的电容器材料.