High frequency embryogenic callus induction and plant regeneration from mature caryposis of big bluestem and little bluestem

Big bluestem (Andropogon gerardii Vitman) and little bluestem [Schizachyrium scoparium (Michaux) Nash.] are native to the North America and are important forage grasses and ornamental grasses. Both grasses are proposed as ideal biomass producers for cellulosic ethanol production. To apply genetic transformation, which is an important tool for incorporating desirable agronomic traits into plants to both species, however requires an efficient and reproducible regeneration protocol. We used mature caryopses from big and little bluestem as explants and tested the effect of various combinations of 2, 4-dichlorophenoxyacetic acid (2, 4-D) (1, 2, 3, 4 or 5 mg l⁻¹) and kinetin (KT) (0, 0.1 or 0.2 mg l⁻¹) on embryogenic callus induction with LS as the basal medium. The highest percentage of embryogenic calli induction occurred on medium containing 2, 4-D alone at 2 mg l⁻¹ for ‘Bison’ and on medium containing 4 mg l⁻¹ 2, 4-D alone for ‘Bonilla’ big bluestem. For little bluestem, the highest percentage of embryogenic callus induction occurred on medium containing 3 mg l⁻¹ 2, 4-D plus 0.1 mg l⁻¹ kinetin, suggesting that addition of KT is beneficial. Shoot regeneration took place on LS basal medium without any plant growth regulator for both species, although the addition of KT increased both regeneration frequency and the number of shoots produced per callus. Rooting of shoots reaching about 2 cm long occurred readily with or without α-naphthaleneacetic acid (NAA). Rooted plantlets were all successfully established in the soil.     

T7 RNA多聚酶基因的克隆、序列测定及其在E.coli中的表达

从BL21(DE3)E．coli菌株中以PCR的方法扩增得到了与T7RNA多聚酶(T7RNApolymerase，T7pol)基因大小一致的DNA片断。将PCR产物纯化后直接克隆到pGEM—T载体中，经酶切鉴定和DNA序列分析表明克隆得到了正确的T7pol基因。将T7pol基因亚克隆入pET-28b( )中，构建得到原核表达质粒pET28T7。该质粒的BL21(DE3)pLysS转化菌在IPTG的诱导下可表达约98800的蛋白，这与T7pol的相对分子质量一致。将该质粒转化DH5α、JMl09、HBl01、BL21(DE3)和BL21(DE3)pLysS等5种不同的宿主菌，仅有转化T7pol酶活性受到抑制的宿主菌BL21(DE3)pLysS才能得到转化子，而其余4种T7T7pol酶活性不受抑制的E．coli宿主菌不能得到转化子。pET28T7原核表达质粒这种仅能在T7pol酶活性受到抑制的宿主菌中才能存活的现象说明本试验所克隆的T7pol基因能正确表达出具有RNA转录酶活性的蛋白。     

马尾松基因库无性系花期观察分析

在福建省沙县官庄林场石景山工区进行马尾松基因库无性系开花习性、花量、花期的观察。观察结果表明：不同无性系着生球花量的差异显著，始花期也有明显的差异，但也存在一定的同步性，大部分无性系雌花开放时间比雄花早４─９天。     

13～18周龄笼养蛋鸭适宜蛋白质和蛋氨酸水平的研究

用 32 4羽满 1 2周龄的金定鸭 ,通过饲养试验、代谢试验、屠宰试验和对试鸭产蛋期 (1 9～ 2 8周龄 )产蛋性能的观察 ,研究了在等能条件下(ME :1 1 51MJ/kg) ,不同蛋白质水平 (CP :1 3 %、 1 5 %、 1 7% )和蛋氨酸水平 (Met/CP :0 0 1 7、 0 0 2 2、 0 0 2 7)组成的 9种饲粮对 1 3～ 1 8周龄笼养蛋鸭的生长性能、养分利用、体成分沉积和产蛋期产蛋性能的影响。结果表明 :该阶段饲粮粗蛋白、蛋氨酸水平对体增重影响不显著 (P >0 0 5) ;体增重与蛋氨酸 (% )呈一定的正相关 (r=0 4574) ;CP为 1 3%、Met/CP为 0 0 2 7组采食量一直较稳定、较高 ,且饲料效率又好。代谢试验 :不同粗蛋白、蛋氨酸水平饲粮对试鸭氮存留率和能量的利用率影响显著(P <0 0 5或P<0 0 1 ) ,且它们随饲粮CP水平的升高而降低 ,随蛋氨酸水平升高而升高 ;磷的利用率也分别以CP为 1 3%的水平和Met/CP为 0 0 2 7的水平极显著地高于其他水平对磷利用率 (P <0 0 1 )。屠宰试验 :当Met/CP为 0 0 2 7时 ,体氮沉积量和体磷沉积量最佳。产蛋性能 :开产日龄和高峰日龄与 1 3～ 1 8周龄CP食入量呈弱的负相关 (r=- 0 1 83 3和r=- 0 0 984) ,与蛋氨酸食入量呈正相关 (r =0 5780和r =0 435 0 )。饲粮粗蛋白、蛋氨酸水平对 1 9～ 2 8周龄     

牛Myostatin基因单核苷酸多态性分析

Myostatin基因即肌肉生长抑制素,是一种肌肉生长的负调控因子。应运PCR-SSCP和测序的方法对中国秦川牛、南阳牛以及国外引入品种皮埃蒙特牛双肌基因的第三外显子进行了多态性分析。结果表明:第三外显子938处G→A的单核苷酸的突变造成了南阳牛、皮埃蒙特牛第三外显子扩增片段多态性,秦川牛则不然。     

真核表达载体构建表达ＰＲＶ闽Ａ株gE基因表位抗原编码区片段的重组质粒

根据伪狂犬病病毒闽Ａ株gE基因表位抗原编码区的序列与身份种真核表达载体pPICZaA、pAcGP67A序列与特性分别设计了两对ＰＣＲ引物。通过ＰＣＲ方法扩增到了两端具有不同酶切位点的gE基因表位抗原编码片段。将这２个片段分别克隆到pPICZaA与pAcGP67A载体，转化大肠杆菌ＴＯＰ１０菌档及ＸＬ１－Ｂlue菌株，获得了含伪狂犬病病毒闽Ａ株gE基因表位抗原编码区的重组质粒pICZaA-FS与pAcGP67A-FS。序测定结果显示两个重组质粒中插入片段的大小与方向均正确。     

Influence of the Indoleacetic Acid-Lysine Synthetase Gene (iaaL) of Pseudomonas syringae subsp. savastanoi on Yield Attributes of Potatoes

The iaaL gene of Pseudomonas syringae subsp. savastanoi encodes an indoleacetic acid-lysine synthetase which conjugates free indoleacetic acid (IAA) with lysine. lAA-lys is biologically less active than free IAA. The iaaL coding region was expressed under the control of the cauliflower mosaic virus 35S promoter and transgenic potato plants were produced (Spena et al. 1991). 35S iaaL potato plants are characterized by increased internodal length and epinastic bending of older leaves. In three greenhouse experiments with plants grown in pots of different size and in two growth chamber experiments tuber number increased in iaaL transgenic plants compared to untransformed and vector-transformed controls of the same genotype. The increase in tuber numbers observed under controlled conditions was reflected in tuber yield which increased in the pot grown transgenics.     

二阶拟线性椭圓型Euler方程组解的Pohozaev型恒等式及其应用

本文对于二阶椭圆型Euler方程组 -∑D_αF_(q_α~i)(x,u,Du)+F_(ui)(x,u,Du)=0得到了一些积分恒等式,利用这些恒等式证明了某些方程组非平凡解的不存在性。     

表达序列标签（ESTs）及其应用

表达序列标签(ESTs)是指从(ESTcDNA文库中随机挑取克隆并对其3’或5’端进行单轮自动测序所获得的短cDNA库列，一般长度为300～500bp。要有效获取ESTs，必须对cDNA文库进行预处理，处理方法有衰减杂交法、均一化法和差异显示法。在dbEST中收录了大量各种生物的ESTs。ESTs广泛应用于鉴定基因、发现新基因、电子克隆、构建遗传学图谱、制备DNA芯片、分子标记、研究基因的差异表达及检验病原微生物等方面。     

Genetic Engineering Approaches to Pig Production*

Modern biotechnology promises a number of new applications in animal breeding and production. Although conventional pig breeding has achieved a high level of efficiency and productivity numerous problems have been encountered with animal health and the loss of meat quality. Selection based on phenotypic performance data of individual animals does not take into account the importance of specific genes and their relevance within a complex regulatory system. In most cases it is therefore difficult to trace back the genetic origins of clinically important disorders. The application of genetic engineering techniques in pig production will facilitate diagnosis, improvement of productivity, and animal health by allowing direct genetic manipulation. Attention must be focussed on the physical and genetic analysis of the procine genome. The isolation and characterisation of genes, DNA-markers, polymorphic DNA-fragments, and their chromosomal assignment will be important prerequisites and tools for the elucidation of genetic disorders. Especially the detection of heterozygous carriers of recessive disorders and their elimination from the breeding stock will increase selection accuracy and decrease the generation intervals. But also the rapid and simple detection of infectious diseases, which is sometimes difficult if not impossible at present, will improve animal health and welfare. Although the production of transgenic animals either by DNA-microinjection into zygotes or the use of embryonal stem cells manipulated in vitro is less straightforward than DNA-based diagnosis it will play an important role in the direct manipulation of the porcine genome and genes. Breeding programmes including the use of transgenic livestock have already been developed. There is no doubt that genetic engineering has reached a degree of practical feasibility, allowing it to play an important role in pig breeding in particular and animal production in general.