Breeding of T. aestivum-Ag. intermedium translocation lines with T. timopheevii cytoplasm and characterization by GISH

A male-sterileT. aestivum-Ag. intermedium partial amphiploid with cytoplasm ofT. timopheevii as a female parent was crossed to common wheat. The hybrid was backcrossed to the male parent several times continually and setf-crossed at last. Two stable lines with common wheat phenotype, H96269-2 and H96278, have been obtained. The chromosome numbers of the two lines are all2n = 42 in somatic cetls. By inoculation test, the two lines show a high levet of resistance to yetlow rust. Through genomicin situ hybridization (GISH) withAg. intermedium total genomic DNA as a probe, it is demonstrated that the two stable lines are all small segmental translocation lines, and the translocated chromosome segments fromAg. intermedium are located on the short arm terminals of wheat chromosomes. Genetics analysis suggests that the yetlow rust resistance gene(s) are probably located on the translocated chromosome segments ofAg. intermedium.     

Physical location of the ricePi-5(t), Glh andRTSV genes by ISH of BAC clones

Mapping of low or single-copy sequences on plant chromosomes has proven difficult because of very low frequency of signal detection. Rice BAC library is being used widely in rice genome research due to its distinctive advantages over other library systems. In this study, two biotin-labeled rice BAC clones closely linked to a rice blast resistance, green leafhopper resistance and tungro spherical virus resistance gene,Pi-5(t), Glh, RTSV, werein situ hybridized to rice chromosomes. They were located on the long arm and short arm of chromosome 4 with FL value of 40% and 100% respectively. The frequency of signal detection reached 46.8% and 59.2%. The signal location were consistent with the selective marker on rice saturated molecular map. The results demonstrated the advantages to locate BAC clones to chromosomes byin situ hybridization and will facilitate the rice low or single-copy gene location by using the BAC library. Supported by the National Natural Science Foundation of China and the Doctorate Vesting Point Foundation of the Education Department of the People's Republic of China Yan Huimin: born in 1964, Lecturer     

Studies inin situ PCR detected by the BrdU antibody technique

Using the BrdU antibody technique followed by an immuno-chemical staining (BAT), the amplification of DNA fragments specific to human Y chromosome on cell specimen slides was efficiently detected. Whether direct BrdU incorporation into PCR products orin situ hybridization with PCR products on slides, the amplified target DNA fragments of specimen were visualized by BAT under the microscope. The availability of BAT and differences in the sensitivity and efficiency between BAT and dig-11-dUTP labeling in cellin situ PCR were discussed. Zhang Xiyuan: born in Oct. 1935, Professor, Current research interest in Cellular and Molecular Biology Supported by the National Natural Science Foundation of China     

Expression of steroidogenic acute regulatory protein and its regulation by interferon-gamma in rat corpus luteum

The steroidogenic acute regulatory protein (StAR) is the key regulatory protein of steroidogenesis.De novo synthesis of StAR protein is required for intramitochondrial translocation of cholesterol to the cytochrome P450 side chain cleavage enzyme which is located on the matrix side of the inner mitochondrial membrane. This is the rate-limiting step of steroid biosynthesis. Usingin situ hybridization and immunohistochemistry we studied StAR expression in various stages of the corpora luteal and its regulation by interferon-gamma (IFNγ) in the adult pseudopregnant rat. The results indicated that expression of StAR in the corpora luteal was correlated with progesteron production and IFNγ was capable of inhibiting its expression.     

Localization and expression of TR3 orphan receptor protein and its mRNA in rat

The cellular localization and specific expression of TR3 mRNA in rat testis were investigated by the method of immuno- histochemistry andin situ hybridization. It was demonstrated that orphan receptor TR3 was expressed in a significant amount in rat testis and TR3 protein and mRNA were specifically localized in germ cells, suggesting that TR3 may have an important function in regulating rat germ cell development.     

Chromosomalin situ hybridization ofCyprinus carpio genomic DNA repetitive sequence CR1

Common carpCyprinus carpio genomic DNA repetitive sequence CR1 has been DIG-labeled and hybridizedin situ against chromosomes of red common carp (Cyprinus carpio L. Xingguo red var. ). It is found that the repetitive sequence CR1 is mainly localized at the centromeric regions of chromosomes of the red common carp. The application of the chromosomalin situ hybridization technique on fish and the relationship between CR1 repetitive sequence distribution and its function have been discussed.     

Expression and regulation of orphan receptor TR2 mRNA in germ cells of cryptorchid testis in rat and rhesus monkey

Expression and cellular localization of orphan receptor TR2 mRNA in relation to germ cell apoptosis in cryptorchid testes of rat and rhesus monkey have been studied by usingin situ hybridization andin situ 3′-end labeling of DNA fragments (TUNEL). The results show that: (i) TR2 mRNA is specifically expressed in the germ cells, mainly in the spermatocytes, round and elongated spermatids. The expression level of TR2 mRNA varies with the seminiferous cycle, (ii) In the rat cryptorchid testes on days 3 and 5 after the surgery, the germ cells began to undergo apoptosis with no evident decrease in TR2 mRNA level. On day 7.5, however, most germ cells underwent apoptosis, while the expression level of TR2 mRNA declined markedly, and TR2 mRNA was rarely expressed on day 10 thereafter, (iii) On days 15 and 20 of the cryptorchid testes of rhesus monkey, TR2 mRNA was only expressed in a few of primary spermatocytes and the mRNA was almost undetectable on days 30, 45, 60. These results suggest that TR2 mRNA probably plays an important role in spermatogenesis and germ cell apoptosis.     

Expression of neurofilament gene in spinal motoneurons during neural regeneration

The techniquein situ hybridization was used to measure the levels of light (NF-L), medium (NF-M) and heavy (NF-H) neurofilament protein subunits mRNA in L_4-6 spinal motoneurons in adult rat during regeneration following a unilateral crush of the sciatic nerve. It was found that the hybridization signals of each neurofilament subunit rnRNA were dramatically decreased in spinal motoneurons postaxotomy by light microscopy. The hybridization signals of NF-L and NF-M mRNA were located in cytoplasm of neurons, whereas NF-H mRNA was found in both nucleus and cytoplasm of neurons. Image analysis showed that the encoding levels of mRNA for each of neurofilament subunit mRNA reduced on the 3rd d and returned to control levels on the 28th d following the lesion. The relative levels of mRNA coding for each neurofilament subunit were significantly different. The lowest level of NF-L mRNA was observed at 5 d postaxotomy, and that of NF-M, NF-H mRNA on the 7th and 10th d after injury. Moreover, the levels of HF-M and NF-H mRNA were reduced much lower and lasted much longer than that of NF-L mRNA. The observations suggest that there were different mechanisms for the regulation of neurofilament subunit genes expression. The reduced neurofilament gene expression may be due to a response to axonal injury and advance the restructure of axonal architecture.     

Follicles in pregnant rat ovary are incapable of steroidogenesis

Antral follicles are present in the ovaries throughout gestation. These follicles are “physically immature” and cannot ovulate under the induction of LH/hCG. The purpose of the present study is to examine whether follicles in pregnant rat ovaries are capable of steroidogenesis. StAR is believed to be the key regulator of steroid hormone biosynthesis. Antisense StAR probe and anti-StAR rabbit serum have been used to detect the StAR expression in ovarian follicles at various stages of pregnant rats. The results indicate that theca-interstitial cells and the membrane granulosa cells at the stages from the estrous or pre-estrous in the normal cycling rat ovary express StAR mRNA and its protein, whereas neither granulosa cells nor the theca cells in pregnant ovary throughout gestation express StAR. These results indicate that the pregnant follicles throughout gestation are incapable of steroidogenesis.     

Rapid diagnosis with FISH for chromosomal abnormality of fetal pyelectasia

Fluorescence in situ Hybridization （FISH） was used to investigate whether the chromosome of the fetus prenatally diagnosed as pyelectasis was normal or not. Amniotic fluid was taken from the pregnant woman whose fetus was detected with pyelectasia by prenatal examination. The chromosome of the amniotic fluid cell without culture was examined with FISH. The result shows that compared with the traditional amniotic fluid cell culture, FISH has the advantages of more rapid, higher sensitivity and specificity, and was 10-12 days earlier to complete the diagnosing than the traditional method. The fetuses detected chromosomal abnormality in each groups were induced during the middle and late trimester, while those fetuses with normal chromosome continued pregnancy, the rate of spontaneous disappearance of pyelectasia decreased as the severity of pyelectasia increased. FISH can satisfy the urgent need in the clinical prenatal diagnosis due to its rapidity to determine whether fetus with pyelectasia was accompanied with chromosomal.     

Breeding and molecular cytogenetic identification of wheat-Thinopyrum intermedium addition lines resistant to powdery mildew

Wheat-related species Th. intermedium was used to cross with common wheat Yannong 15. In the self progenies of the hybrid, two addition lines, Ⅱ-1-7-1 and Ⅱ-3-3-2, stable in cytology, were developed by cytology and powdery mildew resistance identification. Their chromosome number were 2n = 44 and formed 22 bivalents at PMC MI. In F₁ of the two addition lines crossing with Yannong 15, there appeared about one univalent at PMC MI, respectively. Resistance identification in greenhouse and field using the No. 15 and mixed strains of E. gramnis f. sp. tritici showed that they were immune to powdery mildew. Chromosome number and resistance identification using the F² single plants of the addition line crossing with Yannong 15 indicated that the resistant gene was located on the alien chromosomes. In situ hybridization using St and E genomic DNA as probe showed that the added chromosome in the two addition lines probably came from the E genome of Th. intermedium, which indicated that a pair of E genome chromosomes carried a new resistant gene to powdery mildew.     

Localization of orphan receptor TR3 mRNA in early developmental follicles in rat

Byin situ hybridization, the localization of orphan receptor TR3 mRNA has been observed in early developmental follicles. TR3 mRNA is first expressed in the ovarian interstitial cells on day 2 after birth, and then in granulosa cells (GC) in primary follicles on day 4. The expression level of TR3 mRNA in GC increases following the follicular development. Its higher expression can be observed in the outer layer of GC and inner layer of theca cells (TC) on day 6, where the cells present active proliferation and differentiation. The expression of TR3 is in an increasing manner until the large antral follicles on day 30. The mRNA is only expressed in the healthy, but not atretic follicles in adult rat ovaries. Injection of epidermal growth factor (EGF) has dramatically enhanced its expression in the early stage of developmental follicles. It is therefore suggested that TR3 may play a role in regulating growth and differentiation of ovarian somatic cells in the early stage, and its expression is regulated by EGF.     

Preparation of PE/MMT nanocomposite by monomer intercalation and <Emphasis Type="Italic">in situ</Emphasis> copolymerization

The catalyst [(2-ArN=C(Me))₂C₅H₃N]FeCl₂ (Ar = 2-C₆H₄(i-Pr)) with methylaluminoxane (MAO) as cocatalyst was intercalated between layers of montmorillonite (MMT) for ethylene oligomerization. Metallocene catalyst Me₂Si(Ind)₂ZrCl₂ and MAO was then added to form a dual functional catalytic system. A PE/MMT nanocomposite was prepared by copolymerization of ethylene with α-olefins produced in situ from ethylene over the dual functional catalytic system. The catalytic system was of high polymeric activity. The resultant PE/MMT nanocomposites were stable and got increases in tensile strength and temperature of maximum weight loss (T_onset).     

应用基因组原位杂交鉴定杂交后代中的簇毛麦染色体

用地高辛（Digoxigenin-11-dTup)标记的簇毛麦染色体组DNA为探针，以普通小麦“中国春”总DNA作封阻进行基因组荧光原位杂交，对小麦和簇毛麦杂交和回交后代进行检测。结果显示，在杂交后代的28条染色体中有7条簇毛麦染色体，在发生部分染色体加倍的回交代后鉴定出含7条簇毛麦染色体重的易位系。GISH的准确鉴定以及易位系的获得为向小麦导入簇志麦的有用基因提供了宝贵材料。     

Localization and possible role of membrane type metallo-proteinase and tissue inhibitors of metalloproteinase-1 in early stages of placentation

Human placental tissues from the first and second trimesters of gestation have been investigated using riboprobein situ hybridisation of mRNA sequences coding for membrane type metalloproteinase (MT-1-MMP) and tissue inhibitors of metalloproteinase-1 (TIMP-1). Results show that (i) both mRNAs express at a relatively high level in the chorion laeve trophoblast cells and the adjacent decidual cells of fetal membrane; (ii) the most abundant expression of the two mRNAs was found in the extravillous trophoblast between Rohrs and Nitabuch striae of basal plate, trophoblast shell and gland cells of the decidua; (iii) isolated or small groups of cytotrophoblast cells in the chorionic villi and in the cells lining arterioles in decidua and stem villi also expressed both MT-1-MMP and TIMP-1 at defferent extents. The data suggest that the coordinated expression of the MT-MMP and its inhibitor TIMP in defferent cells of the placental tissue may play an essential role in trophoblast invasion and angiogenesis related to placentation in the first two trimesters of gestation. They may also have an ability to effect separation of fetal from material tissue at a favorable junctional site during parturition.     

Analysis of DNA methylation variation in wheat genetic background after alien chromatin introduction based on methylation-sensitive amplification polymorphism

During the process of alien germplasm introduced into wheat genome by chromosome engineering, extensive genetic variations of genome structure and gene expression in recipient could be induced. In this study, we performed GISH （genome in situ hybridization） and AFLP （amplified fragment length polymorphism） on wheat-rye chromosome translocation lines and their parents to detect the identity in genomic structure of different translocation lines. The results showed that the genome primary structure variations were not obviously detected in different translocation lines except the same 1RS chromosome translocation. Methylation sensitive amplification polymorphism （MSAP） analyses on genomic DNA showed that the ratios of fully-methylated sites were significantly increased in translocation lines （CN12, 20.15%; CN17, 20.91%; CN18, 22.42%）, but the ratios of hemimethylated sites were significantly lowered （CN12, 21.41%; CN17, 23.43%; CN18, 22.42%）, whereas 16.37% were fully-methylated and 25.44% were hemimethylated in case of their wheat parent. Twenty-nine classes of methylation patterns were identified in a comparative assay of cytosine methylation patterns between wheat-rye translocation lines and their wheat parent, including 13 hypermethylation patterns （33.74%）, 9 demethylation patterns （22.76%） and 7 uncertain patterns （4.07%）. In further sequence analysis, the alterations of methylation pattern affected both repetitive DNA sequences, such as retrotransposons and tandem repetitive sequences, and low-copy DNA.     

夏季长江口—东海陆架大中型浮游生物分布特征及影响因素

根据2019年夏季长江口及邻近海域17个站位的大中型浮游生物与水质参数分析,研究了大中型浮游生物的空间分布特征及其主要影响因素。在长江口咸淡水混合影响下,长江口-东海陆架断面浮游生物物种数及生物多样性指数均随盐度增加而增大,且淡水及低盐度站位与高盐度站位的优势物种存在显著差异。长江口-东海陆架关键断面13个站位的大中型浮游生物密度及生物量与叶绿素a无显著相关性,与营养盐含量呈显著负相关,表明上行效应并非控制该断面浮游生物分布特征的主要因素。在咸淡水混合所致多因素变化的复杂影响下,该断面大中型浮游生物分布的主要影响因素呈现出盐度相关的区域性差异。首次利用浮游生物在线成像和分类设备（CPICS）在多个高盐度站位开展原位观测,获得了多种大中型浮游生物的原位影像,可对夜光藻、双生水母等大型浮游动物鉴定至种,对桡足类鉴定至亚纲。CPICS垂直布放原位观测与野外垂直拖网采样分析结果所反映的物种数变化趋势具有一致性。