Preparation and analysis of spermatocyte meiotic pachytene bivalents of pigs for gene mapping

Well-spread meiotic pachytene bivalents were obtained by using the prolonged hypotonic treatment com-bined with high chloroform Carnory‘s fixative solution from cells of the testes of domestic pigs. Comparisonin the division index and length of pachytenc bivalents with metaphase chromosomes showed that those ofthe former are 5 times higher and 3.42(1.87-5.98) times longer than those of the latter. Comparative studieson chromomere maps of bivalents and mitotic chromosomal G-bands were conducted by using the chromo-some 12 as a example. Sex vesicle and various shapes of synaptic sex chromosomes have been observed.Two-color PRimed IN Situ (PRINS) labeling has been conducted successfully on pachytene bivalents of pigs.     

Preparation and analysis of spermatocyte meiotic pachytene bivalents of pigs for gene mapping

Well-spread meiotic pachytene bivalents were obtained by using the prolonged hypotonic treatment combined with high chloroform Carnory's fixative solution from cells of the testes of domestic pigs. Comparison in the division index and length of pachytene bivalents with metaphase chromosomes showed that those of the former are 5 times higher and 3.42(1.87-5.98) times longer than those of the latter. Comparative studies on chromomere maps of bivalents and mitotic chromosomal G-bands were conducted by using the chromosome 12 as a example. Sex vesicle and various shapes of synaptic sex chromosomes have been observed. Two-color PRimed IN Situ (PRINS) labeling has been conducted successfully on pachytene bivalents of pigs.     

Preparation and analysis of spermatocyte meiotic pachytene bivalents of pigs for gene mapping

Well-spread meiotic pachytene bivalents were obtained by using the prolonged hypotonic treatment com-bined with high chloroform Carnory’s fixative solution from cells of the testes of domestic pigs. Comparison in the division index and length of pachytene bivalents with metaphase chromosomes showed that those of the former are 5 times higher and 3.42(1.87-5.98) times longer than those of the latter. Comparative studies on chromomere maps of bivalents and mitotic chromosomal G-bands were conducted by using the chromo-some 12 as a example. Sex vesicle and various shapes of synaptic sex chromosomes have been observed.Two-color PRimed IN Situ (PRINS) labeling has been conducted successfully on pachytene bivalents of pigs.     

Meiotic Studies of Diploid Perennial Teosinte and Its Hybrids with Maize

1. Open-pollinated diploid perennial teosinte Microsporocytes at pachytene stage from six different diploid perennial teosinte plants were investigated. It was found that they all had 10 pairs of chromosomes (2n = 20). All of the knobs and large chromomere were terminal (Plate Ⅰ,). The size of the knobs varied from medium to small. The short arms of chromosomes 1 and 2 and the long arms of chromosomes 2 and 3 had a small knob. A large chromomere     

Expression of betaine aldehyde dehydrogenase gene and salinity tolerance in rice transgenic plants

Betaine as one of osmolytes plays an important role in osmoregulation of most high plants. Betaine aldehyde dehydrogenase C BADH) is the second enzyme involved in betaine biosynthesis. The BADH gene from a halophite, Atriplex hortensis, was transformed into rice cultivars by bombarment method. Totally 192 transgenic rice plants were obtained and most of them had higher salt tolerance than controls. Among transgenic plants transplanted in the saline pool containing 0.5% NaCl in a greenhouse, 22 survived, 13 of which set seeds, and the frequency of seed setting was very low, only 10% . But the controls could not grow under the same condition. The results of BADH ac-tivity assay and Northern blot showed that the BADH gene was integrated into chromosomes of transgenic plants and expressed.     

Development of Triticum aestivum-Leymus racemosus translocation lines using gametocidal chromosomes

Specific chromosomes of certain Aegilops species introduced into wheat genome background may often facilitate chromosome breakage and refusion, and finally result in a variety of chromosome restructuring. Such a phenomenon is commonly called gametocidal effect of the chromosomes. The chromosome 2C of Ae. cylindrica is one of such chromosomes. In the present study, scab resistant wheat-L. racemosus addition lines involving chromosomes Lr.2 and Lr.7 were crossed to wheat-Ae. cylindrica disomic addition line Add2C. Then F₁ hybrids were subsequently backcrossed with wheat cv “Chinese Spring”. BC₁ plants with chromosome structural aberration were identified by C-banding. In the self-pollinated progenies of these plants, three translocation lines were developed and characterized by mitotic and meiotic analysis combined with C-banding and fluorescent in situ hybridization (FISH) using biotin-labeled genomic DNA of L. racemosus as probe. Some other putative translocation lines to be further characterized were also found. The practicability and efficiency of the translocation between wheat and alien chromosomes induced by gametocidal chromosomes, as well as the potential use of the developed alien translocation lines were also discussed.     

Induction of small-segment-translocation between wheat and rye chromosomes

A new approach to produce wheat-rye translocation, based on the genetic instability caused by monosomic addition of rye chromosome in wheat, is described. 1 283 plants from the selfed progenies of monosomic addition lines with single chromosome of inbred rye line R12 and complete chromosome complement of wheat cultivar Mianyang 11 were cytologically analyzed on a plant-by-plant basis by the improved C-banding technique. 63 of the plants, with 2n = 42, were found containing wheat-rye translocation or substitution, with a frequency of 4. 91% . Compared with the wheat parent, other 32 plants with 2n = 42 exhibited obvious phenotypic variation, but their com-ponent of rye chromosome could not be detected using the C-banding technique. In situ hybridization with a biotin-la-beled DNA probe was used to detect rye chromatin and to determine the insertion sites of rye segments in the wheat chromosomes. In 20 out of the 32 variant wheat plants, small segments of rye chromosomes were found being inserted into dif     

Inducement of chromosome translocation with small alien segments by irradiating mature female gametes of the whole arm translocation line

Haynaldia villosa Schur. (syn. Dasypyrum villosum Candargy, 2n=14, VV) has been proved to be an important genetic resource for wheat improvement. The development of translocation with small alien chromosome segments, especially interstitial translocation, will be helpful for better utilization of its useful genes. Up to now, most of the reported Triticum aestivum – H. villosa translocation lines are involved in a whole arm or large alien fragments. In this paper, we report a highly efficient approach for the creation of small chromosome segment translocation lines. Before flowering, the female gametes of wheat-H. villosa 6VS/6AL translocation line were irradiated by 60CO-γ ray at 160 Rad/M dosage rate and three dosages (1600, 1920, 2240 Rad). Anthers were removed from the irradiated florets on the same day and the florets were pollinated with normal fresh pollens of T. aestivum cv. Chinese Spring after 2-3 days. Genomic in situ hybridization (GISH) at mitosis metaphase of root-tip cell of M1 plants was used to detect the chromosome structural changes involving 6VS of H. villosa. Among the 534 M1 plants screened, 97 plants contained small segment chromosome structural changes of 6VS, including 80 interstitial translocation chromosomes, 57 terminal translocation chromosomes and 55 deletion chromosomes. For the 2240 Rad dosage treatment, the inducement frequencies of interstitial translo-cation, terminal translocation and deletion were 21.02%, 14.01%, and 14.65%, respectively, which were much higher than those previously reported. The M2 seeds were obtained by backcrossing of 74 M1 plants involving 146 chromosomes structural changes of 6VS, and it was found that the structural aberrations in the M1 plants could be transmitted to their progenies. Irradiating mature female gametes of whole arm translocation is a new and highly efficient approach for creation of small segment chromosome struc-tural changes, especially for interstitial translocations.     

家猪减数分裂粗线期二价体与有丝分裂中期染色体的比较研究

以性成熟公猪睾丸和外周血为材料,采用长低渗、高氯仿卡诺固定液固定和外周血细胞培养制备减数分裂粗线期二价体和有丝分裂中期染色体,通过对二价体和有丝分裂中期染色体分裂指数和长度的比较研究,发现二价体的分裂指数和长度分别是有丝分裂中期染色体的5倍和3.42倍(1.87～5.98);同时以12号染色体为例, 比较了二价体上的染色粒结构带与有丝分裂中期染色体G-带,表明染色粒结构带比中期染色体G-带带纹丰富,而与早中期G-带带纹吻合。 Abstract:Meiotic pachytene bivalents were obtained from porcine testes using prolonged hypotonic treatment combined with high chloroform Carnory's fixative solution. Mitotic metaphase chromosomes were prepared from blood cell culture. Comparative studies on division index and length of pachytene bivalents and mitosis metaphase chromosomes showed that those of the former are 5 times higher and 3.42(1.87～5.98) times longer than the latter, respectively. Chromomere maps of bivalents are more abundant than mitotic metaphase G-bands, while they are correspondent with mitotic early-metaphase G-bands. The result was found by using the chromosome 12 as a sample.     

Effects of crop development on the emission of volatiles in leaves of Lycopersicon esculentum and its inhibitory activity to Botrytis cinerea and Fusarium oxysporum

Volatiles emitted from the leaves of Lycopersicon esculentum at the two-, ten-leaf and anthesis periods were collected by a gas absorbing method and analyzed by gas chromatography （GC）-mass spectrometry. In total, 33 compounds of volaUles emitted from three developmental stage plants were separated and identified, and quantitatively analyzed by the internal standard addition method. All of the samples of volatile were found to be rich in monoterpenes and sesquiterpenes.β-phellandrene and caryophyllene predominated in the volatiles of the leaves of plants at the two- and ten-leaf stages. Furthermore, （E）-2-hexenal were the dominant components in the volatiles emitted from anthesis plants. The results of volatiles analyzed show that the compositions varied depending on the developmental stages. The volatiles emitted from crushed tomato leaves of plants at the anthesis stage had the most strongly inhibitory activity against the spore germination and hyphal growth of Botrytis cinerea and Fusarium oxysporum, followed by ten- and two-leaf plants. However, the activity of volatiles, emitted from the leaves of plants at the two-leaf stage, in inhibiting F. oxysporum was greater than B. cinerea.     

植物染色体G-带的初步研究

本文首次报道了川百台(Lilium davidii)、华山松(Pinus armardii)和七叶一枝花(Paris polyphylla)等植物染色体G-带研究结果。本试验的G-带与以往的C-带不同,C-带每条染色体上一般只有1-4条带,多分布在着丝点附近,而G-带则多达几十条,分布在整条染色体上,带纹清晰,前期染色体带呈颗粒状,中期染色体呈明显的带状,与哺乳动物染色体G-带很相似。G-带的数目取决于染色体浓缩的程度。前期染色体带纹数目是中期的三倍,接近人类高分辨带水平。对G-带带纹采用了自动光谱分析,波峰数值与带纹相符。作者同时介绍了胰酶法在植物染色体G-带中的应用。认为此方法既适合动物亦适用于植物。但植物G-带显示的关键可能不在胰酶法本身,而在合适的分裂时期及染色体处理技术。     

植物染色体高分辨G-带技术研究

本文对植物染色体高分辨 G-带技术进行了比较系统的研究,并首次运用改良的尿素法在野生一粒小麦、玉米、蚕豆、吊兰、川百合等多种植物上诱导出 G-带,带纹清晰,数目多,分布在染色体全长上。前期染色体带呈颗粒状,中期染色体呈明显带状,与哺乳动物染色体 G-带很相似。G-带的数目取决于染色体浓缩程度,中期染色体一条深带到晚前期可显示出2.67条亚带。作者同时比较了胰酶法与尿素法的显带效果。认为两种方法显示的带纹基本相同,尿素法比胰酶法作用温和,显带时间长达数分钟,易于掌握,重复性高,具有更高的应用价值。     

Why plant chromosomes do not show G-bands

Summary Giemsa techniques have refused to reveal G-banding patterns in plant chromosomes. Whatever has been differentially stained so far in plant chromosomes by various techniques represents constitutive heterochromatin (redefined in this paper). Patterns of this type must not be confused with the G-banding patterns of higher vertebrates which reveal an additional chromosome segmentation beyond that due to constitutive heterochromatin. The absence of G-bands in plants is explained as follows: 1) Plant chromosomes in metaphase contain much more DNA than G-banding vertebrate chromosomes of comparable length. At such a high degree of contraction vertebrate chromosomes too would not show G-bands, simply for optical reasons. 2) The striking correspondence of pachytene chromomeres and mitotic G-bands in higher vertebrates suggests that pachytene chromomeres are G-band equivalents, and that this may also be the case in plants. G-banded vertebrate chromosomes are on the average only 2.3 times shorter in mitosis than in pachytene; the chromomeric pattern therefore still can be shown. In contrast, plant chromosomes are approximately 10 times shorter at mitotic metaphase; their pachytene-like arrangement of chromomeres is therefore no longer demonstrable.     

Studies on G-bands and Macrocoils in Plant Chromosomes

Four different methods including trypsin urea, SDS and NaOH are presented for the in situ induction of G-bands and macrocoils on the chromosomes of Secale cereale, Hordeum vulgare and Vicia faba. The bands obtained were numerous and along the whole chromosome, the number of the G-bands was much interrelated with the condensation of chromosomes. The bands of homologous chromosomes in some cells were matchable. The G-banded chromosomes in late prophase have nearly reached high resolution level. When incubation periods were beyond critical time for G-banding, macrocoils were often revealed. Gyre number changed with chromosome condensation and the direction of coils has showed different patterns. Transformation of G-bands into macrocoils was first reported in plant chromosomes. Some chromosomes showing G-bands under light microscope appeared spiral patterns under scanning electron microscope. In this paper the relationship between G-bands and macrocoils in plant chromosomes is also discussed.     

玉米染色体G-带ASG法显带的研究

两个自交系的根尖染邑体经ASG法处理显出了G-带。王米G-带沿整个染色体长轴分布,是一些密切邻近的多重带纹。无论有丝分裂的晚前期、早中期或中期染色体都有这类带纹。每一对同源染色体的两成员G-带带型基本相似,不同染色体或同一染色体的不同区域带纹具有一定的差异。ASG处理前用α-溴萘或放线菌素D预处理都可显出G-带。本文讨论了玉米G-带与哺乳动物G-带的相似点以及用ASG法进行玉米G-带显带应注意的技术问题。     

High resolution bands in maize chromosomes by G-banding methods

Summary It was demonstrated that G-bands are unequivocally present in plant chromosomes, in contrast to what had been formerly believed by plant cytologists. Maize chromosomes prepared by an enzymatic maceration method and treated with trypsin or SDS showed clear G-bands spreading along the chromosomes. The most critical point during the G-banding procedures was the post-fixation with glutaraldehyde solution. Banding patterns were processed by using the chromosome image analyzing system and a clearer image was obtained. Gbanding technique and the image manipulation method described here can be applied to many plant species, and would contribute new information in the field of plant cytology and genetics.     

Induction of G-bands in Root Tip Chromosomes of the Maize

Authors tried to induce G-bands of chromosomes of root-tips in maize (Zey mays L.. everty Sturt) with a variety of technological modifications. The following techniques were found to be more satisfactory: primary root-tips were treated with an aqueous actinomycin D(AMD) solution (70 μg/ml) at room temperature; air dry slides of chromosomes in maize made the chromosomes spread well and plasma off; and then the preparations of the chromosomes were treated with modified methods of Seabright[7] and Utakoji[8] and the technique of aceto-orcin stain. The G-bands of chromosomes of corn were induced with the three methods above. They were shown in Plate 1, 2, 3 and 4 which are similar to the G-bands of chromosomes in human and mammal, and these bands are more consistent in each chromosomes.     

水稻染色体标本制备的风油精法

水稻的染色体较小,不同的染色体在形态上较难区分。常规的压片技术由于很难使染色体分散,且也不能完全排除细胞质的干扰,因而很不适用于水稻染色体核型分析及显带。Kurata 等(1978)采用酶解与火焰干燥技术制备水稻染色体标本,获得清晰的染色体图象,从而成功地进行了水稻染色体的核型分析。陈瑞阳等(1982)参照人类染色体     

Harlequin banding and localisation of sister-chromatid exchanges.

Harlequin banding (HB) was standardised on Indian muntjac chromosomes by superimposing harlequin staining or sister-chromatid differentiation and G-banding after incorporation of bromodeoxyuridine (BrdU) or cholorodeoxyuridine (CldU), and after treatment with BrdU plus mitomycin C (MMC). SCEs were localized on these chromosomes with the aid of the G-band map. There were more SCEs in G-bands than in R-bands in BrdU-incorporated chromosomes. CldU-incorporated chromosomes, however, did not show a preferential localization of SCEs in either G- or R-bands. When BrdU + MMC-induced SCEs were localized in harlequin-banded chromosomes, there was a significantly greater number of SCEs in R-bands; and there was a concomitant reduction in the frequency of SCEs in G-bands, as compared to the SCEs observed in this region after BrdU incorporation alone. Centromeric regions of chromosomes 1 and X had preferred sites for occurrence of SCEs in BrdU-incorporated chromosomes, the preferred sites being more in G-bands after BrdU and CldU incorporation and in R-bands after treatment of BrdU-incorporated chromosomes with MMC. Thus the formation of SCEs is not restricted by structure per se as defined by euchromatin or heterochromatin, but depends on the site of lesion production, type of lesion and repair pathway followed.     

Electron microscopy of G-banded human mitotic chromosomes

Trypsin-treated human metaphase chromosomes stained with Giemsa and uranyl acetate showed clear, reproducible band structures under the transmission electron microscope (TEM). The banding pattern observed with TEM corresponded very closely to the G-band pattern visualized by light microscopy. The TEM images were used for karyotype analyses. Trypsin-treated chromosomes stained with uranyl acetate alone also showed clear G-bands under TEM. Shadow casting in addition to uranyl acetate staining revealed more structural detail of the chromosomes. Chromosome fibers, 200 Å–300 Å in diameter, were observed in the interband regions. Most chromosomes showed the major G-bands under the higher TEM magnification wit0out any trypsin treatment.