Dietary Sterol Preference in the Silkworm,Bombyx mori

Since insects are unable to biosynthesize sterols de novo, sterols must be obtained from dietary sources. Although it has been reported that β-sitosterol is crucial for larval growth in the silkworm, Bombyx mori, little has been investigated concerning the dietary selection of sterols by Bombyx larvae. Here, we demonstrate that Bombyx larvae have the following sterol preference: β-sitosterol >> ergosterol > cholesterol = stigmasterol. Interestingly, Bombyx larvae preferred ergosterol, an inhibitory sterol on larval growth, indicating that sterol selection following first contact of the diet with the mouthpart might be different from the sterol recognition mechanism present in sterol metabolism.     

家蚕细胞遗传学及其应用

由于家蚕染色体数目较多、着丝粒弥散, 在较长时期内, 家蚕染色体识别、核型分析、染色体结构和功能的研究都受到很大限制。近年来, 应用比较基因组杂交、基因组原位杂交、基于细菌人工染色体克隆的原位杂交技术建立了家蚕的细胞学图, 综合分子连锁图构建高密度的细胞遗传学图已成为可能。分子细胞遗传学的应用正在推动家蚕染色体结构和功能的研究, 揭示出家蚕W染色体密集地分布着嵌套结构的逆转座子, 染色体端粒由重复序列(TTAGG)n以及端粒特异的非长末端逆转座子TRAS1和SART1组成, TRAS1、SART1具有较高的转录活性, 可能与维持染色体的稳定性有关。     

Microtubules During Blastoderm Formation in the Egg of the Silkworm, Bombyx mori L.

Microtubules in the silkworm egg, Bombyx mori , were observed by electron microscopy, in order to investigate the relationship between cytoskeletal organelles and the migration of energids, the cleavage nuclei accompanied by the associated cytoplasm, near the egg surface or during blastoderm formation. Numerous microtubules were observed in the associated cytoplasm of an energid even in the interphase of mitosis.
At about 8.5 hr after oviposition, when many energids had already cleft and distributed near the peripheral yolk granule region, long microtubules distributed radially from the perinuclear region to the periphery in the associated cytoplasm. When an energid was protruding, the microtubules above the nucleus distributed at a more acute angle than those under the nucleus. When a blastoderm cell had just been formed, the microtubules were observed only under the nucleus.
Colchicine, an inhibitor of microtubules, stopped the migration of energids and inhibited the formation of blastoderm cells even after many energids had already distributed at the peripheral yolk granule region. The relationship between the microtubules and the migration of energids near the egg surface or during blastoderm formation was discussed.     

Application of Multi-vitamins as Supplementary Nutrients on Biological and Economical Characteristics of Silkworm Bombyx mori L

In order to investigate the effects of supplementary nutrients on silkworm, Bombyx mori, an experiment was conducted with multi-vitamins treatments. Leaves enriched with multi-vitamins (1, 2.5 and 5%) were fed once a day to 4th and 5th instar silkworm larva. The supplementation of the leaves was done by spraying the treatments on them. These treatments resulted in a significant increase in biological and economical parameters such as larval weight, female and male cocoon weight, pupal weight and egg productivity when compared with normal control. Multi-vitamins of 2.5% treatment increased the larval weight by 10.2%, which had the most weight enhancement in the fifth day of 5th instar larvae. However, 5% concentration of multi-vitamins decreased some biological characteristics. These treatments resulted in 4.7% increase of cocoon's weight. In most of the moths the number and weight of eggs had significant difference compared to control, but these supplementary materials caused relatively less egg hatching.     

应用离子注入技术对家蚕形态学及遗传学进行的初步研究(英文)

离子注入技术是将某种元素的原子进行电离,并使其在电场中加速,在获得较高的速度后射入固体材料表面。在离子注入过程中,被电离的离子在电场作用下加速运动,离子靠着本身获得的动能进入基体表面,在表层中运动的离子与基体原子作用损失能量后在一定的位置停留下来。该技术自６０年代问世以来,主要用于材料改性等方面。８０年代中期,我国学者开始将其用于农作物育种方面的研究,大大拓宽了离子注入技术的应用领域。所用实验材料的基因及表现型见Ｔａｂ３,我们将氢离子（Ｅ＝３５ＭｅＶ）注入处于胚胎发育后期的家蚕卵内（Ｔａｂ１）,观察其对家蚕形态及遗传方面的影响,结果表明：（１）在家蚕胚胎发育的已４期注入氢离子,其半致死剂量ＬＤ５０为１ｘ１０１０～１ｘ１０１１ｃｍ２这一区间之内;当剂量达到１ｘ１０１２ｃｍ２时,已全部致死（Ｆｉｇ．１＆Ｔａｂ．２）;（２）注入氢离子能够使家蚕在第１腹节上产生褐斑（Ｆｉｇ．２）的频率增高。并首次观察到因注入氢离子而导致家蚕出现非成对的褐斑（Ｆｉｇ．３＆Ｔａｂ．４）。（３）在氢离子注入剂量为１ｘ１０１０ｃｍ２时,能够诱变产生大量的嵌合体家蚕,并且诱变频率高达３８．５％（Ｆｉｇ．４＆Ｔａｂ．５）,这样高的     

Isolation and Identification of a Pathogen of Silkworm Bombyx mori

A pathogenic bacterial strain, ST-1, was isolated from a naturally infected silkworm. The strain was identified on the basis of its physiological and biochemical properties and the results of sequence analysis of its 16S rRNA gene. The results of the 16S rRNA gene sequence analysis revealed that ST-1 shared the highest sequence identity (more than 99%) with Pseudomonas chlororaphis subsp. aurantiaca. ST-1 bacteria were gram-negative and 0.7-0.9 × 1.3-1.5 μm long, short rods with rounded ends. The strain could utilize sodium citrate, malonate, D-glucose, sucrose, D-fructose, D-mannose, and L-arabinose. Pathogenicity of ST-1 for silkworm could be depicted as a linear regression of the logarithm (y) of ST-1 concentration against probability (x) (y = 0.4040 + 0.0600x). The median lethal concentration (LC(50)) was 2.12 × 10(4) cfu/ml. In conclusion, ST-1 was identified as Ps. chlororaphis subsp. aurantiaca. This is the first report that Ps. aurantiaca is a pathogen for silkworm Bombyx mori.     

Effects of BmCPV Infection on Silkworm Bombyx mori Intestinal Bacteria

The gut microbiota has a crucial role in the growth, development and environmental adaptation in the host insect. The objective of our work was to investigate the microbiota of the healthy silkworm Bombyx mori gut and changes after the infection of B. mori cypovirus (BmCPV). Intestinal contents of the infected and healthy larvae of B. mori of fifth instar were collected at 24, 72 and 144 h post infection with BmCPV. The gut bacteria were analyzed by pyrosequencing of the 16S rRNA gene. 147(135) and 113(103) genera were found in the gut content of the healthy control female (male) larvae and BmCPV-infected female (male) larvae, respectively. In general, the microbial communities in the gut content of healthy larvae were dominated by Enterococcus, Delftia, Pelomonas, Ralstonia and Staphylococcus, however the abundance change of each genus was depended on the developmental stage and gender. Microbial diversity reached minimum at 144 h of fifth instar larvae. The abundance of Enterococcus in the females was substantially lower and the abundance of Delftia, Aurantimonas and Staphylococcus was substantially higher compared to the males. Bacterial diversity in the intestinal contents decreased after post infection with BmCPV, whereas the abundance of both Enterococcus and Staphylococcus which belongs to Gram-positive were increased. Therefore, our findings suggested that observed changes in relative abundance was related to the immune response of silkworm to BmCPV infection. Relevance analysis of plenty of the predominant genera showed the abundance of the Enterococcus genus was in negative correlation with the abundance of the most predominant genera. These results provided insight into the relationship between the gut microbiota and development of the BmCPV-infected silkworm.     

The Effect of Level and Source of Dietary Selenium Supplementation on Eggshell Quality

A 16-week-long experiment was performed to compare the effect of sodium selenite (SS) and selenium-enriched yeast (SY) supplementation on eggshell quality and also to evaluate breaking force correlation with other parameters of shell quality originating from hens fed with selenium supplementation. One hundred Shaver 579 hens (27 weeks old) with similar body size were randomly divided for five dietary treatments: basal diet without selenium supplementation and basal diets with two levels of selenium supplementation (0.4 or 0.8 mg/kg) via SS or SY. No adverse effect of Se inclusion in hen's feed, regardless of its source, on shell breaking force, shell deformation, shape index, shell thickness and shell percentage, were observed throughout the current study (P > 0.05). Moderate correlations were found between breaking force and nondestructive shell deformation for all diets (P < 0.05). There was no significant overall correlation between egg breaking force and shell thickness or/and percentage shell in the presence of selenium supplemention (P > 0.05). Shape index in all four selenium-supplemented groups was not related to the breaking force (P > 0.05). Selenium supplementation of up to 0.8 mg/kg, regardless of its source, in the diet of laying hens in their first phase of laying does not adversely affect eggshell quality.     

Effect of Carotenoid Depletion on the Hatching Rhythm of the Silkworm, Bombyx mori

To investigate the functional involvement of carotenoid in the insect circadian rhythm, we observed the effect of carotenoid depletion on the hatching patterns of the silkworm under several light conditions. When eggs were transferred from continuous light (LL) to continuous darkness (DD), an overt hatching rhythm was initiated. The first hatching peak, which was observed at 13.2 h after the transfer in the carotenoid-depleted silkworm, was reduced remarkably in both control groups (carotenoid-rich and carotenoid-depleted but with lutein supplementation), though subsequent peaks occurred with similar timing. Under LD 4 : 20, LD 12 : 12 and LD 20 :4, hatching patterns depended on the dietary carotenoid and the light intensity of the photophase. At a low light intensity, carotenoid depletion suppressed hatching as a masking effect just after light-on. Under LD 4 : 20 at a low light illumination, hatchings in the carotenoid-depleted silkworm were observed during scotophase, but few larvae hatched during scotophase in the control groups. Our findings suggest that carotenoid is not indispensable for the photoreception, formation and entrainment of the circadian hatching rhythm, but that a carotenoid-dependent process that is induced by light-off and damps out in an hour-glass mode, suppresses the hatching during darkness without affecting the circadian oscillation. The possibility that this carotenoid-dependent process is involved in the photoperiodic induction in the silkworm was discussed.     

新霉素抗性基因在家蚕中的插入和表达

构建含新霉素抗性基因（ｎｅｏｍｙｃｉｎｒｅｓｉｓｔａｎｃｅｇｅｎｅ,ｎｅｏＲ）的重组质粒ｐＦＮ,经ＨｉｎｄＩＩ酶切后,用基因枪将ＤＮＡ片段导入家蚕早期受精卵中（Ｇ０代）。孵化的Ｇ１、Ｇ２代蚁蚕均经含新霉素的人工饲料添食２４ｈ后,筛选出新霉素抗性的个体（能正常生长发育的）改为桑叶饲养。于Ｇ２代的５龄第二天从后部丝腺抽提总ＤＮＡ,再以ｎｅｏＲ的ｃＤＮＡ为探针进行Ｓｏｕｔｈｅｒｎ杂交检测。结果表明ｎｅｏＲ基因已转入家蚕ＤＮＡ中,获得了含ｎｅｏＲ的转基因蚕。     

Scanning Electron microscopic Study on the Early Development of Silkworm Eggs (Bombyx mori L.)

Morphological changes of the surface of eggs of the silkworm Bombyx mori L. were studied during early developmental stages by scanning electron microscopy. The egg surface was covered with numerous microvilli at least until 4 h after oviposition. At 6 h the microvilli were replaced by ruffle-like microprojections. This suggests that developmental changes of the surface structure may occur without direct influence of cleavage nuclei. Immediately before blastoderm cell formation, microvilli reappeared in the presumptive groove area. The ruffles seen on the apical portion of newly-formed blastoderm cells gradually became flattened, while microvilli developed on the lateral side of the cells. The mode of blastoderm cell formation is different from the typical one seen in most species of insects.     

Molecular Markers (RAPD) Associated with Growth, Yield, and Origin of the Silkworm, Bombyx mori L. in India

To identify the molecular markers associated with growth and yield parameters in silkworm, Bombyx mori, RAPD profiles generated with seven UBC primers for fourteen silkworm stocks, originated from China, Japan, India, and Russia, were statistically analyzed. Stepwise multiple regression analysis establishes significant association of 45 markers with larval span, growth indices and four cocoon yield parameters relevant for silk production and t-test attest significance of the association of 89.5_{1500 bp} and 54.13_{300 bp}, respectively, with longer larval duration and high cocoon weight. The validity of this selection of markers was further supported with discriminant function analysis (DFA) done on the basis of Mahalanobis D ² statistics. The two indices of yield/growth were also tested with DFA, which helped in identifying a few markers and thereby opened scope of using such marker (e.g., 91.11_{900 bp}) for incorporating molecular markers in the breeding program for crop improvement in silkworm.     

差异蛋白质组学在家蚕中的研究进展

差异蛋白质组是蛋白质组学的一个重要分支,通过对蛋白质组表达谱的比较,揭示细胞生理或病理状态的进程与本质,发现具有关键作用的蛋白。近年来,家蚕差异蛋白质组学发展迅速且涉及面广,已然成为家蚕蛋白质组学研究的热点。对差异蛋白质组学的主要研究方法,及在家蚕中的研究进展做一简要评述。     

蜕皮激素对家蚕卵黄原蛋白基因表达的调控

蜕皮激素（20-hydroxy ecdysone,20E）是由前胸腺分泌的能调节节肢动物昆虫纲、甲壳纲等动物蜕皮的激素. 家蚕属于完全变态昆虫,一生经历卵、幼虫、蛹、成虫四个发育时期. 20E在家蚕发育过程中有不可替代的作用. 本研究通过酶联免疫吸附反应（ELISA）测定家蚕变态期前后血淋巴中20E滴度,发现在卵黄原蛋白基因 (Bombyx mori vitellogenin, BmVg) 高量表达前的雌雄血淋巴中20E含量没有明显差异,但含量变化趋势与BmVg表达趋势相一致. 用20E注射上蔟后60 h的家蚕和添食5龄家蚕, 能诱导雌性和雄性脂肪体中BmVg表达和蛋白合成. 调查处理后的家蚕所产卵中的卵黄磷蛋白（BmVn）含量及重量,发现蚕卵中BmVn含量及蚕卵重量都明显增加|本研究还通过脂肪体体外培养,证明了20E是直接作用于脂肪体来诱导BmVg的表达. 本研究结果表明,20E是通过诱导脂肪体中BmVg的转录来增加蚕卵中BmVn积累,从而最终使得蚕卵重量也相应增加.     

Isolation and Characterization of Prothoracicotropic Hormone from Silkworm,Bombyx mori

Members of the small family of Arabidopsis PSEUDO-RESPONSE REGULATORS (PRR1/TOC1, PRR3, PRR5, PRR7, and PRR9) play roles close to the circadian clock in Arabidopsis thaliana. We have reported that the rice (Oryza sativa) genome also encodes a set of PRR counterparts (designated OsPRR1, OsPRR37, OsPRR59, OsPRR73, and OsPRR95 respectively). To gain new insight into the molecular functions of OsPRRs, we carried out genetic complementation analyses by introducing two representative rice genes, OsPRR1 and OsPRR37, into the corresponding Arabidopsis loss-of-function mutants (toc1 and prr7 respectively). The results showed that these OsPRR and AtPRR genes are genetically interchangeable at least in part, suggesting the conserved clock-associated function of these OsPRRs.     

家蚕化性及其变化的调控

化性是家蚕的一种重要性状,不仅受遗传基因控制,还受温度、光线等外界环境条件的影响而发生变化。介绍了家蚕化性变化的因素及其机理,并根据家蚕化性的特性,论述了养蚕生产上调控家蚕化性以提高丝量丝质、防止生种与再生卵发生的技术措施。     

Studies on the Sterol of Bombyx mori L

In whole stages of Bombyx silkworm it was shown with gas-liquid chromatographic systems that silkworm-sterols consist of three sterols such as cholesterol, β-sitosterol, and campesterol, at least, and the sterols in the 5th instar larva contain afore-mentioned three sterols and an additional unknown sterol. Moreover, the sterol composition of the silkworm was studied in various stages.