Detection of a highly heterozygous locus in recombinant inbred lines of rice and its possible involvement in heterosis

Forty-seven recombinant inbred (RI) lines derived from a cross between two indica rices, cv Phalguna and the Assam land race ARC 6650, were subjected to restriction fragment length polymorphism (RFLP) analysis using cloned probes defining 150 single-copy loci uniformly dispersed on the 12 chromosomes of rice. Of the probes tested, 47 detected polymorphism between the parents. Heterozygosity was calculated for each line and for each of the polymorphic loci. Average heterozygosity per line was 9.6% but was excessive (>20%) in the 5 lines that seemed to have undergone outcrossing immediately prior to harvest. Average heterozygosity detected by each probe across the 47 RI lines was 9.7%. The majority of probes revealed the low level of heterozygosity (<8%) expected for F₅-F₆ lines in a species showing about 5% outbreeding. On the other hand, 7 probes exhibited heterozygosity in excess of 15%, while with a eighth probe (RG2 from chromosome 11) heterozygosity varied according to the restriction enzyme employed, ranging from 2% with SaII to 72% with EcoRV. The presence of 34 recombination sites in a segment of the genome as short as 24 kb indicates a strong selection for recombination between two neighbouring loci, one required as homozygous for the Phalguna allele, and the other heterozygous. Since selection was principally for yield advantage over that of the high-yielding parent, Phalguna, one or both of these loci may be important for heterosis in this cross. The results also indicate that heterozygosity as measured by RFLP can depend on the particular restriction endonuclease employed.     

Effects of Guaianolide derivative I on fibre initiation and elongation in Gossypium arboreum L. in relation to adverse effects of cool temperature

Cotton ovules collected during late September with prevalent night cool temperature (15°C), cultured at 30°C/15°C i.e. cycling temperatures in Beasley and Ting medium had very few epidermal cells showing bulging. Supplementing cultures with guaianolide derivative I (E-13 methyldehydrocostus lactone) promoted fibre initiation. At—1 day preanthesis (DPA), IAA oxidase activity declined in guaianolide-treated cultures but increased during the elongation phase and was enhanced during the secondary wall thickening phase. However, o-diphenol oxidase activity was adversely affected during the fibre initiation phase. The activities of all the other enzymes studied viz. acid invertase, phenylalanine ammonia lyase, -glucosidase and IAA oxidase increased, except -galactosidase, during the later phase in comparison with the controls. The present study indicates that guaianolide derivative I triggers early initiation and promotes fibre elongation by regulatin o-diphenols and IAA-oxidation levels, which in turn check wall loosening. Considerable enhancement in the soluble acid invertase activity by this compound suggests its role in apoplastic sucrose hydrolysis, thereby preventing its accumulation.Abbreviations DPA days pre anthesis - DAC days after culturing - DAA days after anthesis     

小麦提型不育系和杂交种种子生活力遗传改良初探

本试验共选用了4个提型不育系(A系)及其保持系(B系)、4个恢复系(R系)及其川7B/R4份F~1|代材料,用不同遗传背景的B、R系及川7B/R材料与A系杂交,种子成熟时收获干燥考种,度过休眠期后进行发芽试验。结果表明,父本对F~0|种子千粒重存在胚乳直感现象;川3A、川4A与其它B系杂交,其F~0|种子的千粒重、饱满度、发芽率和发芽势均有不同程度的提高;川4A×R和川6A×R的杂种种子千粒重多分别比川4A×川7B/R和川6A×川7B/R的高,但前种组合(A×R)的种子发芽和发芽率远不如后一种组合(A×川7B /R)的种子,且前种组合的穗发芽率也较高。作者认为,利用A系与农艺性状相近、但遗传背景各异的B系杂交,或在R系中输入抗提型细胞质负影响的高种子生活力基因, 是提高A系和杂交种种子生活力的值得注意的途径。 Abstract:The objective of this paper is trying to grope for ways of improving sced viability of A-line and hybrid in wheat with T.timopheevi cytoplasm.Four A lines and their B lines,4 restorers (R line) and 4 crosses of Chuan 7B/R were used.The combinations of A×B,A×R and A×Chuan 7B/R were madc,and 1 000-grain weight (GW),rate of pre-harvest sprouting(RPHS),germinating energy(GE) and germination percentage (GP) of their F0 seeds were investigated.The results showed that the GW,full weight,GE and GP of the seeds of Chuan 3A and 4A×other B lines were higher than those of Chuan 3A×3B and chuan 4A×4B;although the GW of Chuan 4A and 6A×R were heavier than those of Chuan 4A and 6A×Chuan 7B/R,the GE and GP of the latter crosses increased largely and their RPHScs were less.Therefore,it was considered as effective ways for improvement on seed viability,that A lines cross with other B lines having different genetic background but similar agronomic characters and that the gene(s) concerning high seed viability were transferred into restorers.     

Immortalized hypothalamic luteinizing hormone-releasing hormone (LHRH) neurons: A new tool for dissecting the molecular and cellular basis of LHRH physiology

Summary 1. Two LHRH neuronal cell lines were developed by targeted tumorigenesis of LHRH neuronsin vivo. These cell lines (GN and GT-1 cells) represent a homogeneous population of neurons. GT-1 cells have been further subcloned to produce the GT1-1, GT1-3, and GT1-7 cell lines. While considerable information is accumulating about GT-1 cells, very little is currently known about the characteristics and responses of GN cells.2. By both morphological and biochemical criteria, GT-1 cells are clearly neurons. All GT-1 cells immunostain for LHRH and the levels of prohormone, peptide intermediates, and LHRH in the cells and medium are relatively high.3. GT-1 cells biosynthesize, process, and secrete LHRH. Processing of pro-LHRH appears to be very similar to that reported for LHRH neuronsin vivo. At least four enzymes may be involved in processing the prohormone to LHRH.4. LHRH neurons are unique among the neurons of the central nervous system because they arise from the olfactory placode and grow back into the preoptic-anterior hypothalamic region of the brain. Once these neurons reach this location, they send their axons to the median eminence. With respect to the immortalized neurons, GN cells were arrested during their transit to the brain. In contrast, GT-1 cells were able to migrate to the preoptic-anterior hypothalamic region but were unable correctly to target their axons to the median eminence. These problems in migration and targeting appear to be due to expression of the simian virus T-antigen.5. While GT-1 cells are a homogeneous population of neurons, they are amenable to coculture with other types of cells. Coculture experiments currently under way should help not only to reveal some of the molecular and cellular cues that are important for neuronal migration and axonal targeting, but they should also highlight the nature of the cellular interactions which normally occurin situ.6. GT-1 cells spontaneously secrete LHRH in a pusatile manner. The interpulse interval for LHRH from these cells is almost identical to that reported for release of LH and LHRHin vivo. GT-1 cells are interconnected by both gap junctions and synapses. The coordination and synchronization of secretion from these cells could occur through these interconnections, by feedback from LHRH itself, and/or by several different compounds that are secreted by these cells. One such compound is nitric oxide.7. GT-1 cells have Na⁺, K⁺, Ca²⁺, and Cl^– channels. Polymerase chain reaction experiments coupled with Southern blotting and electrophysiological recordings reveal that GT-1 cells contain at least five types of Ca²⁺ channels. R-type Ca²⁺ channels appear to be the most common type of channel and this channel is activated by phorbol esters in the GT-1 cells.8. LHRH is secreted from GT-1 cells in response to norepinephrine, dopamine, histamine, GABA (GABA-A agonists), glutamate, nitric oxide, neuropeptide Y, endothelin, prostaglandin E₂, and activin A. Phorbol esters are very potent stimulators of LHRH secretion. Inhibition of LHRH release occurs in response to LHRH, GABA (GABA-B agonists), prolactin, and glucocorticoids.9. Compared to secretion studies, far fewer agents have been tested for their effects on gene expression. All of the agents which have been tested so far have been found either to repress LHRH gene expression or to have no effect. The agents which have been reported to repress LHRH steady-state mRNA levels include LHRH, prolactin, glucocorticoids, nitric oxide, and phorbol esters. While forskolin stimulates LHRH secretion, it does not appear to have any effect on LHRH mRNA levels.     

The methylene blue colorimetric microassay for determining cell line response to growth factors

The validity of the methylene blue colorimetric microassay for determining the response of monolayers of human ovarian tumour cell lines to different growth factors was investigated. Linearity of the relationship between cell density and optical density was confirmed for each cell line (r=0.989–0.999,p<0.001), and when initial cell density was optimised to give exponential growth over the assay period, differences in response to medium supplements were obvious. The response of target cells to growth factors, obtained using the methylene blue assay, were compared with, and found to parallel, previously documented responses obtained non-colorimetrically. Thus Mink lung epithelial cells (MLEC) were inhibited by TG (Holleyet al., 1983), EGF had an inhibitory effect on A431 cells (Gill & Lazar, 1981; Barnes, 1982), and the mesothelial cell line showed a proliferative response to EGF and hydrocortisone (Connell and Rheinwald, 1983).The methylene blue colorimetric microssay was found to be a simple, reliable, sensitive method with low variability, for determining the response of cultured cells to growth factors.     

A novel approach to the analysis of the initiation of embryo development in gramineae

An in vivo model system to study the initiation of embryo development is presented. From the so-called Salmon system of wheat (alloplasmic lines with a 1BL-1RS chromosome translocation), three completely isogenic and homozygous lines were produced by selection for uniformity in about 20 selfing/backcross generations as well as between sublines of doubled haploids. The line (aestivum)-Salmon is male fertile and sexual. The lines (caudata)-Salmon and (kotschyi)-Salmon are male sterile and have a parthenogenetic capacity of about 90%. The expression of nuclear-cytoplasmic male sterility is different for the two parthenogenetic lines. The initiation of autonomous embryo development at defined developmental stages of the ovaries and the maximum degree of parthenogenesis are identical in both parthenogenetic lines as proved by the auxin test and progeny analyses. The protein patterns from ovary extracts of the three isogenic lines were identical for more than 200 spots of 2-D polyacrylamide gels, confirming their homogeneity. However, one protein (P 115.1) was found 3 days before and during anthesis only in ovaries of the parthenogenetic lines. It seems to be involved in the initiation of parthenogenesis.     

Identification of new products of S-aminoethylcysteine ketimine autoxidation

Summary In continuation of a previous work (Pecci et al., 1993), dedicated to the detection of the autoxidation products of S-aminoethylcysteine ketimine (AECK), we give here data for the identification of 2,3,6,7-tetrahydro-4H-[1,4]thiazino[2,3-b]thiazine, thiomorpholine-3-one and 5,5, 6,6-tetrahydro-2,2-dihydroxy-3,3-bi-2H-thiazine among the products of AECK autoxidation. Identification has been done on the basis of mass spectrometry and NMR spectral analyses of the isolated products.Abbreviations TLC thin layer chromatography - HPLC High performance liquid chromatography - AECK S-aminoethylcysteine ketimine     

Inhibition of replicative DNA synthesis in nuclei of Strongylocentrotus intermedium urchin embryo by 2′,3′-dideoxy-3′-aminonucleoside 5′-triphosphates

2′,3′-Dideoxy-3′-aminonucleoside 5′-triphosphates have been shown to be inhibitors of replicative DNA synthesis in isolated nuclei of sea urchin embryo. These compounds inhibit the Okazaki fragment synthesis. The effect of 2′,3′-dideoxy-3′-aminothymidine 5′-triphosphate and arabinothymidine 5′-triphosphate is reversible when adding the corresponding substrate for DNA synthesis, 2′-deoxythymidine 5′-triphosphate.     

Intracellular distribution of acridine derivatives in platelets and their suitability for cytoplasmic pH measurements

The site and mechanism of accumulation of acridine derivatives into platelets and their isolated organelles were investigated. In addition, their suitability as indicators of cytoplasmic pH was analysed. Direct microscopic observation showed that quinacrine and 9-aminoacridine are concentrated inside organelles in platelets. Using fractionation studies, the acridine derivatives were found to accumulate particularly in dense and α-granules. Uptake into these organelles is driven by a pH differential across their membrane (acidic inside). Because of their cellular distribution, acridine derivatives were found to be poor indicators of cytoplasmic pH. In contrast, a poorly permeant dicarboxylated fluorescein derivative, generated in situ by cytosolic enzymes, is shown to be a more reliable probe of intracellular pH. The results are compared with previous reports of the use of 9-aminoacridine as a cytoplasmic pH probe in platelets and of quinacrine as a selective dense-granule marker.