Cloning and Expressing of a Gene Encoding Cytosolic Copper/Zinc Superoxide Dismutase in the Upland Cotton

In this study, a gene encoding a superoxide dismutase （SOD） was cloned from senescent leaves of cotton （Gossypium hirsutum）, and its expressing profile was analyzed. The gene was cloned by rapid amplification of cDNA ends （RACE） method. Northern blotting was used to show the profile of the gene expression, and the enzyme activity was mensurated by NBT deoxidization method in different growth periods. The full length of a gene of cytosolic copper/zinc superoxide dismutase （Cu/Zn-SOD） was isolated from cotton （GenBank Accession Number： DQ445093）. The sequence of cDNA contained 682 bp, the opening reading frame 456 bp, and encoded polypeptide 152 amino acids with the predicted molecular mass of 15.03 kD and theoretical pI of 6.09. The amino acid sequence was similar with the other plants from 82 to 87%. Southern blotting showed that the gene had different number of copies in different cotton species. Northern blotting suggested that the gene had different expression in different tissues and development stages. The enzyme activity was the highest in peak flowering stage. The cotton cytosolic （Cu/Zn-SOD） had lower copies in the upland cotton. The copper/zinc superoxide dismutase mRNA expressing level showed regular changing in the whole development stages; it was lower in the former stages, higher in latter stages and the highest at the peak flowering stage. The curve of the copper/zinc superoxide dismutase mRNA expressing level was consistent with that of the Cu/Zn-SOD enzyme activity. The copper/zinc superoxide dismutase mRNA expressing levels of different organs showed that the gene was higher in the root, leaf, and lower in the flower.     

Effect of in ovo zinc injection on the embryonic development,tissue zinc contents,antioxidation, and related gene expressions of broiler breeder eggs

Two experiments were conducted to investigate the effect of in ovo zinc(Zn) injection on the embryonic development, tissue Zn contents, antioxidation and related gene expressions of fertilized eggs of Arbor Acres broiler breeders. Experiment 1 was conducted to determine an optimal embryonic age for early in ovo injection. A total of 720 fertilized eggs with similar weights were randomly allotted to 4 treatments with 6 replicates per treatment and 30 eggs per replicate in a completely randomized design. The eggs were injected with 0.1 m L sterilized water at 3, 6 and 9 embryonic days of incubation(E3, E6 and E9) or non-injection(the control), respectively. The results from experiment 1 showed that E3 and E6 injections increased(P0.05) the embryonic mortalities, and decreased(P0.05) hatchabilities compared to the non-injected control, but no differences(P0.05) between E9 injection and the non-injected control were observed in either embryonic mortality or hatchability. The findings suggest that the E9 is the optimal embryonic age for early in ovo injection. In experiment 2, a total of 672 fertilized eggs with similar weights were randomly allocated to 7 treatments with 6 replicates per treatment and 16 eggs per replicate in a completely randomized design. The eggs were injected with 0(the negative control), 50, 100, 150, 200, or 250 μg Zn/egg as reagent grade Zn SO4·7 H2 O in a 0.1-m L solution, or non-injection(the positive control), respectively at E9–10. The results from the experiment 2 demonstrated that no differences(P0.05) among 50 and 100 μg Zn/egg groups and the negative control were observed in the embryonic mortality and hatchability, however, the injection of 200 μg Zn/egg increased(P0.05) the embryonic mortality, and injections of 150 and 200 μg Zn/egg decreased(P0.05) hatchabilities compared with the controls. The embryonic tibia Zn contents at E20 were increased(P0.05) by injections of 150, 200 and 250 μg Zn/egg. Zinc injection did not affect(P0.05) malonaldehyde(MDA) contents, copper-and Zncontaining superoxide dismutase(Cu Zn SOD) activities and m RNA expression levels in the liver and heart of chick embryos at E15 and E20. Compared with the negative control, injections of 50, 150 and 200 μg Zn/egg up-regulated(P0.05) the metallothionein(MT) mR NA expression levels in the embryonic liver at E20. These results indicated that in ovo Zn injections increased Zn contents in the embryonic tibia and MT m RNA expression levels in the embryonic liver at E20, however, injections of 150–200 μg Zn/egg were harmful to the embryonic development.     

Intake and Digestibility of Native and Exotic Grasses Fed Ad libitum to Djallonke Sheep in South Benin

Native （Andropogon gayanus, Panicum maximum and Pennisetum purpureum） and exotic grasses （Brachiaria ruzi：iensis, Panieum maximum cv. C1 and T58） consumed in tropical humid zone were evaluated by estimating their voluntary intake and in vivo digestibility. The measurements were performed with 4 male Djallonke sheep （28 kg LW） fed ad libitum in metabolic cages, during a 10-days in vivo trial, preceded by a 14-days accommodation period. Each grass was studied at 3 stages of development. The samples （distributed grasses, refusals and faeces） were analysed for organic matter （OM）, crude fibre （CF）, neutral detergent fibre （NDF）, acid detergent fibre （ADF）, acid detergent lignin （ADL） and crude protein （CP） contents. Voluntary digestible organic matter intake （VDOMI） and voluntary digestible crude protein intake （VDCPI） were calculated （in g kg ^-1 LW075） as synthetic parameters to compare the grasses. The chemical composition of the distributed forages varied significantly （P 〈 0.001） between species or cultivars and development stages. After 4-5 leaves per tiller stage, CP contents decreased in all species and lowest in native species. Andropogon gayanus was the most l ignified species at any stage of development. At the end of the rainy season, Pennisetum purpureum leaves and Brachiaria ruziziensis had the highest CP. For all the grasses, CP contents were higher （and CF contents lower） in the consumed material, compared to the offered forages, showing thereby the high selectivity by the sheep. Dry matter intake, OM and CP digestibility varied greatly （P 〈 0.001） between forages and development stages with high interactions between these factors （P 〈 0.001）. Voluntary digestible organic matter intake and VDCPI were the highest （P 〈 0.05） for native Panicum maximum and Pennisetum purpureum. There was a relationship between digestibility and intake parameters, and ADL content of the grasses seems to be an important factor determining at least digestibility.     

Analysis of Developmental Proteome at Egg Stage of Drone Honeybees （A. m. ligustica）

An investigation on the proteome of drone egg development of native Italian bee （Apis mellifera ligustica Spinola,1806） was carried out in order to prove up the characteristics in protein expression and regulation at egg stage and open out the molecular mechanism of the development. The experiment was carried out by two-dimensional gel electrophoresis. The results showed that there were 200, 242 and 233 proteins in a wide rang of molecular weight （12.42-169.60 kDa） and in a relatively narrow scope of pI （4.50-9.00） detected on day 1, day 2 and day 3, respectively, during the developmental process of the drone egg. Meanwhile, 164 protein spots were resolved at all the images （i.e., the protein was consistently expressed） along with the egg development, among which 7 were significantly up-expressed （P 〈 0.05） and 4 were significantly down-expressed （P 〈 0.05） while 79 had no significant differences （P 〉 0.05）. In addition, the specific proteins expressing proteins on day 1, day 2 and day 3 were 11, 18 and 18, respectively. Besides, 17 proteins expressed both on day 1 and day 2 but silenced on day 3, and 43 proteins expressed both on day 2 and day 3 but silenced on day 1, while only 8 proteins expressed both on day 1 and day 3 but silenced on day 2. The results indicate that 2-d-old eggs are at the most active expressional stage in the development of drone egg. The protein expressing at all images suggests that it should be indispensable for drone egg development, but their expression pattern is different. The proteins expressing at a specific age of egg suggest that specific proteins are needed in different developmental stages to regulate. And there are more house-keeping proteins in the developmental process of the drone egg than that of worker egg, and it will provide more targets for gene improvement.     

Dynamic Changes in the Calcium Content of Several Apple Cultivars During the Growing Season

Dynamic changes in calcium content were investigated in eight apple cultivars. The results showed that the calcium concentration in leaves and shoots increased with fruit development. The cultivars displayed only a small difference in the calcium concentration during the early stage of development, the difference became very significant at the late stage of development, especially in shoots. In shoots, for example, calcium content was highest in Starkrimson （19 638.6 mg kg^-1） and lowest in Fuji （8 751.3 mg kg^-1）. Calcium concentration was highest in young fruits and was found to decrease with the growth of fruit, and was characterized by a dramatic drop at the rapid expansion stage. There was a significant difference among cultivars. Young Starkrimson fruits contained the highest calcium concentration of 506.52 mg kg^-1 among cultivars tested, followed by Pink Lady and Fuji. The calcium concentration in mature fruits from high to low is as follows： Starkrimson, Sansa, Pink Lady, Senshu, Gala, Fuji, Red General and New Century. In this study, it was found that eight cultivars continuously assimilated calcium during the whole growing season, especially at the young stage when fruit took up 35-46% of total calcium. The calcium content in fruitlets was low in all cultivars; in the expansion stage, there was rapid absorption of about 30% of total calcium, whereas in the ripening fruit, content of calcium was reduced. The calcium accumulation increased with fruit growth in stalk, similar to that in fruit.     

洞庭黄鲶胚胎发育观察(英文)

[Objective] The research aimed to provide technical basis for fry rearing of Silnrus asotus in Dongting Lake.[Method] The induced spawning medicine was used in the experiment to conduct artificial induced spawning and fertilization for obtaining round green fertilized eggs.According to embryonic development,the morphological characteristics of embryo at different developmental stages were recorded detailedly through microscope.[Result] The embryonic development of Silnrus asotus in Dongting Lake was divided into 7 stages,namely, blastoderm stage,cleavage stage, blastula stage, gastrula stage,neurula stage,organogenesis stage and pre-hatching stage.After hatched for 37 h 20 min in water at 22-24 ℃, fries were come out.[Conclusion] The time sequence of Silnrus asotus in Dongting Lake was basically similar to that of other catfish,while its hatching time was shorter than that of other fish in Siluriformes.     

Analysis of the Lignin Contents and Related Enzymes Activities in Seed Coat Between Black-Seeded and Yellow-Seeded Rapes （Brassica napus L.）

One pair of near isonegic yellow/black seeded rape （Brassica napus L） were used as experimental materials to study the changes of lignin contents and enzymes activities of 4-coumarate： CoA ligase （4CL）, Cinnamyl alcohol dehydrogenase （CAD） and ferulate 5-hydroxylase （F5H） in seedcoats during the stage of seed development. The variation analysis showed that the changes of lignin contents and enzyme activities of 4CL, CAD and F5H in seed coat had significant differences between black- and yellow-seeded rapes, and also between different development stages. The correlation analysis demonstrated that the lignin contents were positively correlative to the activities of all three enzymes investigated in the study, and the interactions between them in the seed coat of the two lines. For yellow-seeded rape, the correlation coefficient （0.7262018） of lignin content and the interaction between 4CL and F5H was significant, the lignin contents were highly positively correlative to the activities of CAD and F5H, the interaction between 4CL and CAD, and the interaction between CAD and F5H. For the black-seeded rape, only the lignin content was highly positively correlative to the activity of F5H （the correlation coefficient was 0.772949）, the other correlation coefficients, i. e. lignin contents to 4CL, CAD activities, the interactions between the three enzymes were not significant although all the correlation coefficients were above 0.5000. The results suggested that 4CL, CAD and F5H regulated the biosynthesis of lignin in seedcoat of rapes, leading to the lignin contents in the seedcoats of the yellow-seeded rape much lower than that of the black-seeded line, and affecting the thickness of the developing seedcoats in rapes. Therefore, it was likely to change the seedcoat ratio by overexpressing or suppressing the activities of one of the enzymes, both of them or all of them.     

Conservation of Iris bismarckiana Regel （Iridaceae） Using Plant Regeneration via Somatic Embryogenesis

In an attempt to propagate and conserve the rare, showy bulbous plants of Iris bismarckiana, newly recorded to the flora of Jordan and to contribute to the conservation the wild lris species in Jordan, a simple rapid, time consuming protocol has been developed using plant regeneration via somatic embryogenesis. Somatic embryogenesis was induced in zygotic embryo culture on Murashige and Skoog （MS） medium supplemented with 2,4-dichlorophenoxy acetic acid （8 mg/L） as the sole plant growth regulator, where both embryogenesis calli and somatic embryos were induced. Separation of embryos from embryo clusters was necessary to enhance the frequency of germination. Germination was stimulated by separation of embryos successfully from embryo clusters and transferred onto fresh MS medium. Data obtained were analyzed as a complete random design with three replications. Calli fragments that were transferred to embryogenesis induction medium （EIM） produced white embryo-like globular structures within two weeks. Within three more weeks, clusters of structures at various stages of development could be found on the same callus. The applied technique is rewarding and encouraging for further research on the endangered wild species of Iris in Jordan.     

Nutrient Status and Dynamics of Trifolium spp. at Different Growing Stages

The nutrient status and dynamics of three clover （Trifolium spp.） varieties at different growth stages were investigated with the crude protein content as an indicator of nutrient equilibrium. The crude protein, neutral detergent fiber （NDF）, phosphorus （P）, and calcium （Ca） content followed a decreasing trend at the initial growing stage and increased later. NDF and Ca content peaked in May, while crude protein and P content reached the highest in May and the lowest in July. The crude ash content followed an uncertain pattern as plants growing. The ash content of both wild-type red clover （T. pratensis L.） and ＇Sativum＇ decreased at the beginning and then increased. However, the ash content of ＇Revendef white clover （T. repens L.） decreased all through the experimental period. The general trend was observed for all three clover varieties as the ash content was the highest at vegetative growing stage （May）, and the acidic detergent fiber （ADF） content increased with time. The nutrient equilibrium value was ranked as ＇Revendei＇〉wild-type red clover〉＇Sativum＇.     

Review of History and Recent Development of Organic Farming Worldwide

The history of the organic farming worldwide was reviewed in this paper. The development of the organic farming worldwide had gone through three stages, emergence, expansion, and growth. The contributors and their thoughts during the different development stages of the organic farming were briefly introduced. And the development status of the organic farming worldwide was reviewed from the aspects of land area under organic management, land area under organic management in percentage of total agricultural area, and world markets for organic products. Besides, the main existing problems for the further development of the world＇s organic farming, as well as the development status, problems and strategies of the Chinese organic farming were discussed.     

Myofiber development during embryonic to neonatal development in duck breeds differing in muscle growth rates

Little is known about the muscle developmental patterns during embryonic to neonatal development in ducks.We investigated the developmental patterns in the lateral gastrocnemius muscles of Gaoyou and Jinding ducks differing in their muscle growth rates during the final stages of egg incubation and the first week after hatching.Expression of the MyoD gene was quantified by quantitative real-time PCR(qRT-PCR).The average cross-sectional area and diameter of the fibers increased from embryonic day 21(E21),peaking at E27,and then declining slightly 7 d after hatching.The density of the fibers decreased initially but increased after hatching in both breeds and sexes.The within-breed variation in muscle fiber-type composition was greater than the average variation between the breeds.Overall,the percentage of type Ⅰ fibers increased and that of type lIb fibers decreased consistently.However,the percentage of type lla fibers was almost constant as development proceeded in both duck breeds.The profiles of MyoD mRNA expression were similar in both breeds,and a significantly positive relationship was observed between the expression of MyoD and the percentage of type lIb fibers.This study firstly revealed the characteristics of duck muscle development and differences between the two breeds differing in growth rates.Moreover,type lIb fibers might convert to type Ⅰ fibers in the lateral gastrocnemius,while MyoD may potentially function in controlling the muscle fiber phenotype during the secondary myogenesis of muscle development.     

Profile Analysis of the Proteome of the Egg of the High Royal Jelly Producing Bees （Apis mellifera L.）

The protein composition of the egg development in the high royal jelly producing bees （Apis mellifera L.） was investigated. This pioneer study was to separate and quantify the proteins in the egg of the high royal jelly producing worker bees （Apis mellifera L.） by using two-dimensional gel electrophoresis along with their three-day development. The results showed that 160, 195, and 176 proteins, with a wide range of molecular weight （17-80 KDa） and relatively narrow scope of pI （4. 00-8.40） could be detected on day 1, day 2, and day 3, respectively, during the developmental process of the egg. Meanwhile 44 protein spots were constantly detected along with the egg development. Among them 36% were in the uptrend along with the egg development, 14% were in the downtrend, and 39% were of the largest expressed volume on day 2. In addition, the specific proteins were expressed on day 1, day 2, and day 3 （89, 77, and 80, respectively）. Besides the coexistent and specific proteins, 24 proteins were expressed on day 1 and day 2, but silenced on day 3, 49 proteins were expressed on day 2 and day 3, but silenced on day 1, only 3 proteins were expressed on day 1 and day 3, but silenced on day 2. The result indicates that egg development is a sequential and complex gene controlled process, where the eggs of day 2 express the most active proteins. The coexistent proteins suggest that it is conservative and indispensable for this event. These specific proteins suggest that the different developmental stage needs specific proteins to regulate it.     

Effect of increasing levels of nitrogen on yield of tea in different agro-ecological conditions of Pakistan

Field experiment was conducted on mature tea bushes of Qi-Men variety at two different altitudes from sea level i.e. at National Tea Research Institute Shinkiari （1000 m） and on farmer＇s field in Konsh valley at village Battal （1500 m）. Different levels of nitrogen with constant dose of phosphorus and potassium were applied to observe the optimal dose of nutrients for highest yield under local edaphic conditions. The experiment was repeated for three consecutive years. The highest yield of fresh tea leaves were obtained by the application of 420 kg ha^-1 N followed by the application of 360 kg ha^-1 N at 1000 masl. The highest dose of nitrogen also reduced significantly the soil pH in the upper （0-15cm） layer depth during 1998 and 1999. During the year 2000, the reduction in soil pH was observed by the application of highest dose of nitrogen but was non-significant at 5% level. The lower soil depth （15-30 cm） showed the reduction in pH with the highest dose of nitrogen during the experimental year but was statistically non significant. At the higher altitude of 1500 masl the highest yield of fresh tea leaves was obtained by the application of 420 kg ha^-1 during the experimental years but the increase was significant during the year 1999 and 2000. The yield of the fresh tea leaves was positively correlated with the amount of nitrogen application at both the altitudes. The increasing amount of nitrogen decreased the soil pH at both the depths of 0-15cm and 15-30cm. Unlike the lower altitude （1000 m） there was no any significant reduction in the soil pH during 1998 and 1999. A significant reduction in soil pH was found in the year 2000.     

Growth Performance,Carcass Yield and Meat Quality of Korean Native Duck

This study was carried out to investigate the growth performance, carcass yield and meat quality of large-type Korean native ducks （KND）. A total of 420 day-old ducklings were divided into four treatments （seven replicates/treatment, 15 birds/replicate） by strains （A and B） and gender （male and female） in a 2 × 2 factorial arrangement under complete randomized design. The body weight （BW） of female KND was higher at week 2 and for male it was higher in week 8 （P 〈 0.05）. BW gain, feed intake and feed conversion ratio of male KND were higher than those of female KND （P 〈 0.05） during week 0 to week 8. The carcass yield of both strains increased from 6 weeks to 8 weeks （P 〈 0.05）. The breast meat ratio was the highest at week 8 whereas the leg meat ratio was the highest at 6 weeks of age in both strains （P 〈 0.01）. Lightness （L＊）, yellowness （b＊） and juiciness of meat were the highest at 6 weeks （P 〈 0.05）. Moisture, ash and fat contents of meat were the highest at 7 weeks （P 〈 0.05） and protein content was the highest at 8 weeks （P 〈 0.01）. These results may provide the basic data on growth performance and meat quality of large-type KND, which can be used for genetic improvements and rearing at commercial scale.     

Phytic Acid Contents of Wheat Flours from Different Mill Streams

The phytic acid contents of eight fractions of wheat flours from different mill streams and those in wheat brans, which were separated by different sieves into various sizes, were determined and analyzed. A rapid method for phytic acid assay by adding thioglycolic acid （mercapto acetic acid） with 2,2-bipyridine was used, with an acidic iron-Ⅲ-solution of known iron content. The amount ofphytate was indicated by the decrease in iron in the supernatant. Significant differences were observed in phytic acid content among different milling streams and different cultivars （P〈0.05）. On an average, coarse bran had the highest phytic acid content （53.85 mg g^-1）, and the shorts had 28.48 mg g^-1. The B5 break flour had a higher phytic acid content （4.8 mg g^-1） than the B7 （2.75 mg g^-1） and B8 （2.03 mg g^-1） reduction flours. Lower values were found in the B3, B6 and B7 flours （1.07, 0.79, and 0.76 mg g^-1, respectively）. The phytic acid contents of bran decreased with smaller bran sizes, ranging from 54 to 5.09 mg g^-1.     

Differential Gene Expression Between Hybrids and Their Parents During the Four Crucial Stages of Cotton Growth and Development

The study aims to clarify the differential gene expression between cotton hybrids and their parents in order to better understand the molecular basis of cotton heterosis. The research focused on cotton heterotic and lower heterotic hybrids and their parents during the four crucial stages, which were analyzed using a differential display technique. The results indicated that there were both quantitative and qualitative differences in gene expression amongst them. The quantitative differences include over- and under-expression of parental genes and the dominant expression of highly-expressed parental genes in hybrids. In contrast, the qualitative differences are the following： （i） Bands were observed in both parents but not in the F1 hybrid （BPnF1）; （ii） bands occurred in either of the parents but not in the F1 hybrid （UPnF1）; （iii） bands presented only in the F1 hybrid but not in either of the parents （UF1nP）; and （iv） bands were detected in either of the parents and the F1 hybrid （UPF1）. Overall, the major differences of gene expression occurred in the qualitative level and four related differential patterns were observed. Furthermore, the amount of differential patterns during the flowering stage was relatively higher than those of other stages. At this juncture, both the amount of hybrid-specific expression patterns at flowering stage and the silenced expression patterns at boll-forming stage in highly heterotic hybrids were found higher than those in the lower heterotic ones. It was concluded that significant differences of gene expression in leaves were present between cotton hybrid and its parents during the whole growing stages. Hence, these differences might be responsible for the observed cotton heterosis.     

Effects of Drying Processes on the Antioxidant Properties in Sweet Potatoes

The effects of different drying methods （hot-air drying, microwave drying and vacuum-freeze drying） on the antioxidant activity and antioxidants in sweet potato （lpomoea batatas L. Lain.） tubers were investigated to determine the potential drying process. Sweet potato tubers were cut into 5 mm thick slices, steamed at 100℃ for 10 rain, then dried in either hotair, microwave, or vacuum-freeze. The dried sweet potatoes in microwave possessed the highest antioxidant activity, while the lowest activity was observed in hot-air dried samples. The phenolic contents were positively correlated with scavenging activity and reducing power of DPPH^＋. The microwave drying retains the highest antioxidant activity with the highest content of phenolic compounds in dried sweet potatoes. β-carotene and ascorbic acid showed minor contribution to the antioxidant activity in dried sweet potatoes.     

安康地区泥鳅胚胎发育的观察

[目的]了解安康地区泥鳅的胚胎发育全过程.[方法]用DOM和LHRH-A2对泥鳅进行催产,将所产卵子进行人工授精后所获的胚胎24～26℃、pH值7.0～7.5的淡水中培养,进行泥鳅的胚胎发育观察,详细描述从受精卵到初孵仔鱼期的27个具体发育时期的形态特征和发育时间.[结果]泥鳅胚胎发育可划分为卵裂期、囊胚期、原肠胚期、神经胚期和器官形成期.在24～26℃、pH值7.0～7.5的淡水中,尼鳅胚肥从受精到孵化出膜历时30 h 45 min.[结论]为泥鳅的人工繁殖和遗传育种研究提供了重要的参考. Abstract： [Objective] Aim to know the whole process of embryonic development of loach.[Method]DOM + LHRH-A2 was used to induce spawning of loach,then after fertilization,the embryos were cultured into freshwater water with temperature from 24 to 26 ℃ and pH from 7.0 to 7.5.The embryonic development of loach was observed and 27 concrete morphological characteristics and development time of loach from fertilized egg to newly hatched larval period were described in detail. [Result] The embryonic development of loach could be divided into cleavage stage,blastocyst stage,gastrula stage,neurula stage and organogenesis stage.The loach embryo from fertilized egg to out membrane period was 30 h 45 min in fresh water from 24 to 26 ℃ and pH from 7.0 to 7.5. [Conclusion] it provided important reference for studying artificial propagation and genetic breeding of loach.