Studies on the Genus Paeonia (1)—Report of Karyotypes of Some Wild Species in China

In the present paper 8 species with 15 populations of the genus Paeonia L. (if P. papaveracea and P. japonica are recognised as species) were collected from Sichuan, Shaanxi and Hebei provinces (see the Appendix for detail of the materials). The micrographs of their somatic metaphase (also Mii in the case of P.veitchii) are shown in Plates 1-4, the karyotype formulae, ranges of chromosome length and classification of karyotypes according to Stebbins (1971) are shown in Table 5: the idiograms in Figs. 1-2, and the parameters of chromosomes in Table 1-4. The essential points are mentioned as follows: (1) Chromosomes of the various species in the section Modan have so far been examined and they are all diploid, the two species in the section Onaepia are also diploid, and thus tetraploids exist only in the section Paeonia. (2) Chromosomes in the genus Paeonia are relatively stable except for the differentiation of ploidy. The karyotypes (Table 1-4) show no differences among different taxa in Sect. Modan and the same can also be said about the taxa in Sect. Paeonia (Table 1). Not only are the karyotypes very similar, but also among the members within either section have the same parameters of chromosomes, and, differences, if occur, are not statistically significant. Between the two sections, however, the situation is different. The arm ratios of the first pairs of chromosomes in Sect. Modan are 1.53, 1.52 and 1.48 (Table 1), but those in Sect. Paeonia are 1.12-1.28 (Table 2-4), 95% confidence limits are 1.46-1.60 for the section Modan and 1.07-1.28 (1.21-1.35 only for PB85078) for the section Paeonia, not overlapping, which indicates that the two sections have differentiated in respect of the first pairs of chromosomes. (3)The population PB85024, which belongs to the P. obovata complex, has a karyotype of 2B (stebbins, 1971), which is a new one in the genus Paeonia. This karyotype is a stable one, for several individuals in the population are uniform in this respect, which shows that Stebbins’ (1971) generalization that all the species in Paeonia have 2A does not hold true. (4) Three populations of P. obovata complex studied in this work from Sichuan and Shaanxi are all tetraploids, and one from Hebei is a diploid. From the present work and the previcus reports, the materials from Japan and Korea, no matter whether flowers are pink or white, are diploids, those from Heilongjiang Province (with both pink and white flowers) (Liu Ming-yuan, personal communication) and from Heibei Province (with pink flowers) in China are also diploids, the one from Sakhalin (pink flowers) is tetraploid, those from Priamur of the Soviet Union are a tetraploid (with white flowers) and a diploid (with pink flowers), and those from Shaanxi (the Qinling Range) and western Sichuan (with both pink and white flowers) are all tetraploids. As far as we have now known, ploidy in this particular complex is correlated with the geographical distribution: diploids are found in the central part, tetraploids occur in the northern limits, and in the south letraploids are the only cytotype. (5) The materials of P. mairei from western Sichuan and Shaanxi (the Qinling Range) are found all to be tetraploids, which shows that two cytotypes, diploid and tetraploid, exist in this species, but the geographical distribution pattern of these two cytotypes is to be revealed in the future investigation.     

The Genetics of Dopa Decarboxylase in DROSOPHILA MELANOGASTER I. Isolation and Characterization of Deficiencies That Delete the Dopa-Decarboxylase-Dosage-Sensitive Region and the α-Methyl-Dopa-Hypersensitive Locus

A detailed cytogenetic investigation of 16 overlapping deficiencies in the 36C-40A region on the left arm of the second chromosome (2L) in Drosophila melanogaster is reported. These deficiencies permit a localization of both the dopa-decarboxylase-dosage-sensitive region and the α-methyl-dopa-hypersensitive locus, l(2)amd, to the same region, 37B10-37C7.     

Studies on sex-organ development. Isolation and characterization of an oestrogen receptor from chick Müllerian duct.

An oestradiol-binding macromolecule was observed in the left Müllerian duct of the 15-day female chick embryo. The embryonic receptor binds oestradiol with a high affinity and low capacity, having a Kd of 3.2 X 10(-9)M and a maximal number of sites of 5.45 fmol/10(6) cells in the left Müllerian duct. The receptor is protein in nature, as suggested by its susceptibility to proteolysis; in addition, it is organ- and steroid-specific. Judging by glycerol-gradient analysis, the hormone receptors in the cytosol are present in 8S and 4.5S forms, and the 8S form could be dissociated into a 4.5S form in the presence of 0.5M-KCl. A 4.5-6S receptor could be extracted from the nuclei. Under physiological salt conditions, the embryonic receptors bind to DNA-cellulose and can be eluted when the salt concentration is increased to 0.5M-KCl. Determination by isoelectric focusing indicates that the isoelectric point is 5.8 for the 8S and 6.9 for the 4.5S receptor.     

14. Studies on Poliomyelitis Virus—Concentration and Purification of the Virus

ABSTRACT

For the purpose of determining the immunogenic potency of polio virus, relatively large amounts of concentrated virus material were prepared which had titres of the order of 10¹⁰ T.C.I.D.jo per ml. These were obtained by pervaporating large quantities of tissue culture fluid containing approximately 10⁶⁵ T.C.I.D.JQ per ml.     

Characterization of Glyoxalase I (E. coli)—Inhibitor Interactions by Electrospray Time-of-Flight Mass Spectrometry and Enzyme Kinetic Analysis

Potential inhibitors of the enzyme glyoxalase I from Escherichia coli have been evaluated using a combination of electrospray mass spectrometry and conventional kinetic analysis. An 11-membered library of potential inhibitors included a glutathione analogue resembling the transition-state intermediate in the glyoxalase I catalysis, several alkyl-glutathione, and one flavonoid. The E. coli glyoxalase I quaternary structure was found to be predominantly dimeric, as is the homologous human glyoxalase I. Binding studies by electrospray revealed that inhibitors bind exclusively to the dimeric form of glyoxalase I. Two specific binding sites were observed per dimer. The transition-state analogue was found to have the highest binding affinity, followed by a newly identified inhibitor; S-{2-[3-hexyloxybenzoyl]-vinyl}glutathione. Kinetic analysis confirmed that the order of affinity established by mass spectrometry could be correlated to inhibitory effects on the enzymatic reaction. This study shows that selective inhibitors may exist for the E. coli homologue of the glyoxalase I enzyme.     

Fructose-1,6-bisphosphatase in mantle of the sea mussel Mytilus galloprovincialis Lmk—I. Purification and ion requirements

• 1.1. The enzyme fructose-1,6-bisphosphatase was purified from the mantle of the sea mussel Mytilus galloprovincialis Lmk. The purified enzyme showed a single band in SDS-polyacrylamide gel electrophoresis. The mol. wt and subunit mol. wt of the enzyme were 105,000 and 27,000, respectively.
• 2.2. Divalent cations are essential for the enzyme activity. In the absence of chelating agents, FBPase 1 exhibits hyperbolic kinetics with respect to Mn²⁺, Zn²⁺ and Mg²⁺. The K_m for Mg²⁺ is lower than the physiological concentration of cation in the tissue, whereas its K_m for Mn²⁺ and Zn²⁺ is greater than the respective in vivo concentrations.
• 3.3. The joint action of Mg²⁺ and Zn²⁺ increases the affinity of the enzyme for the substrate Fru-1,6-P₂, though V_max is reduced.
• 4.4. Na⁺ strongly inhibits the enzyme even at very low concentrations. K⁺ has no effect whatsoever.

     

Studies on the Chinese Woodsiaceae (3)—Phylogeny and Speciation

The history of the study on the woodsiodes is briefly surveyed in the paper. The family Woodsiaceae is recognized by the present author. The relationships among the species and the probable evolution of the family are discussed based on author's cytological and comparative morphological studies, and are indicated by Wagner's method, with numerical values as the indices. Woodsiaceae may have originated from the common ancestor of modern Dicranopteris and Sticherus of Gleicheniaceae, and evolved from it into two main branches, i.e., Woodsia and Protowoodsia. The origin of species through polyploid series are discussed, and W. andersonii, W. subcordata, W. alpina and Cheilanthopsis indosiosa considered as fertile allopolyploids; the probable way of speciation is also suggested for these species.     

Studies on β-glucanases. Some properties of a bacterial endo-β-(1→3)-glucanase system

A commercial enzyme preparation, originally obtained from a Flavobacterium(Cytophaga), was fractionated by continuous electrophoresis, giving a protein fraction which hydrolysed laminarin, carboxymethylpachyman, barley β-glucan, lichenin and cellodextrin in random fashion. This enzymic activity was not very stable. Ion-exchange chromatography and molecular-sieve chromatography on Bio-Gel P-60 showed that this activity was due to two specific β-glucanases, an endo-β-(1→3)-glucanase and an endo-β-(1→4)-glucanase. The two enzymes occur in both high- and low-molecular-weight forms, the latter endo-β-(1→3)-glucanase having a molecular weight of about 16000.     

A Comparison Studies on Ultrastructure and Calcium Distribution in Fertile and Sterile Anthers of a Genic Male-sterile Rapeseed （Brassica napus L.）

It‘s common understanding that plant male sterility is closely related to cell ultrastructure or cell microstructure,plant physiological and biochemical metabolism during the generation and development of anthers. The materials used for the study were fertile and sterile anthers in various stages of a genic male-sterile rapeseed     

Purification and Some Properties of Acid-stable α-Amylases from Shochu koji (Aspergillus kawachii)

Aspergillus kawachii α-amylase [EC 3.2.1.1] I and II were purified from shochu koji extract by DEAE Bio-Gel A ion exchange chromatography, Sephacryl S-300 gel chromatography (pH 3.6), coamino dodecyl agarose column chromatography and Sephacryl S-200 gel chromatography. By gel chromatography on a Sephacryl S-300 column, the molecular weights of the purified α-amylase I and II were estimated to be 104,000 and 66,000, respectively. The isoelectric points of α-amylase I and II were 4.25 and 4.20, respectively. The optimal pH range of α-amylase I was 4.0 to 5.0, and the optimum pH of α-amylase II was 5.0. The optimum temperatures of both α-amylases were around 70°C at pH 5.0. Both α-amylases were stable from pH 2.5 to 6.0 and up to 55°C, retaining more than 90% of the original activities. Heavy metal ions such as Hg^{2 +} and Pb^{2 +} were potent inhibitors for both α-amylases.     

Some foliicolous ectoparasites and associated fungi from Jabalpur (M.P.) — I

Summary The present paper describes nine ectoparasitic foliicolous fungi from Jabalpur (M.P.) India. These include four new species viz.Asterina woodfordiae Sahni onWoodfordia fruticosa (L.)Kurz.,Schiffnerula fici Sahni, onFicus infectoria Roxb.,Sarcihella fumosus Sahni onAegle marmelos Corr. andSarcinella odinae Sahni onOdina wodier Roxb.,Acremoniella sarcinellae Pat. &Har.,Fumago vagans Pers.,Stigmella palawanensis Syd. andSchiffnerula cassiae are new fungus records for this country.Mitteriella zizyphina Syd. has been recorded onZizyphus xylopyra Willd., for the first time from this state.Z. xylopyra is a new host record forM. zizyphina.     

Structural and functional importance of theβ-turn in proteins. Studies on proline-containing peptides

We report here two sets of results on proline-containing linear peptides, one of which brings out the role of theβ-turn conformation in the structure of nascent collagen while the other points to the functional importance of the β-turn in calcium-binding proteins. Based on the data on peptides containing the -Pro-Gly-sequence, we had proposed and experimentally verified that theβ-turn conformation in these peptides is a structural requirement for the enzymic hydroxylation of the proline residues in the nascent (unhydroxylated) procollagen molecule. Our recent data, presented here, on the conformation of peptides containing both the -Pro-Gly- and -Gly-Pro-sequences reveal that while theβ-turn in the substrate molecule is required at the catalytic site of prolyl hydroxylase, the polyproline-II structure is necessary for effective binding at the active site of the enzyme. Thus, peptides containing either theβ-turn or the polyproline-II structure alone are found to act only as inhibitors while those with the polyproline-II followed byβ-turn serve as substrates of the enzyme. In another study, we have synthesized the two linear peptides: Boc-Pro-D-Ala-Ala-NHCH₃ and Boc-Pro-Gly-Ala-NHCH₃ each of which adopts, in solution, a structure with two consecutiveβ-turns, as judged from circular dichroism, infrared and nuclear magnetic resonance data. Drastic spectral changes are seen in these peptides on binding to Ca²⁺. Both the peptides show a distinct specificity to Ca²⁺ over Mg²⁺, Na⁺ and Li₊. A conformational change in the peptides occurs on Ca²⁺ binding which brings together the carbonyl groups to coordinate with the metal ion. These results imply a functional role for theβ-turn in Ca²⁺ — binding proteins.     

Effect of 2,4-D on Seedling Physiology and Cytogenetical Studies in Triticum aestivum and Phalaris minor (Gramineae)

Effects of 2, 4 -D on seedling growth and chromosomal abnormalities were studied in Triticum aestivum and Phalaris minor. Seeds were soaked at different concentrations of 2,4 -D （0.01%, 0.1%, 1.0% ） for 4, 8, 12 and 16 hours. 2,4-D suppressed the germination more severely in P. minor than in T. aestivum. Shoot and root length was retarded with the increase of concentration and time of treatment in both species. Generally radical was more negatively affected than coleoptile and emergence of radical was not observed at 1.0% concentration at 8, 12, and 16 hours of treatment in T. aestivum while in P. minor there was a total lack of radical emergence at 1.0% concentration for all durations of treatment. Stiff and curled roots and undifferentiated callus like scutellar tissues were observed in T. aestivum, while in P. minor the coleoptile obtained was lean, pale green in colour and was lying flat on filter paper. Mitotic index decreased, while chromosomal abnormalities, bridges and laggards were increased with the increase of concentration and soaking time however, laggards were not observed in T. aestivum. Clumping and chain formation of chromosomes at metaphase was also noticed in P. minor.     

Effect of 2,4-D on Seedling Physiology and Cytogenetical Studies in Triticum aextivum and Phalaris minor(Gramineae)

Effects of 2, 4 -D on seedling growth and chromosomal abnormalities were studied in Triticum aestivum and Phalaris minor. Seeds were soaked at different concentrations of 2, 4 -D (0.01%, 0.1%, 1.0%) for 4, 8, 12 and 16 hours. 2, 4 -D suppressed the germination more severely in P. minor than in T. aestivum. Shoot and root length was retarded with the increase of concentration and time of treatment in both species. Generally radical was more negatively affected than coleoptile and emergence of radical was not observed at 1.0% concentration at 8, 12, and 16 hours of treatment in T. aestivum while in P. minor there was a total lack of radical emergence at 1.0% concentration for all durations of treatment. Stiff and curled roots and undifferentiated callus like scutellar tissues were observed in T. aestivum, while in P. minor the coleoptile obtained was lean, pale green in colour and was lying flat on filter paper. Mitotic index decreased, while chromosomal abnormalities, bridges and laggards were increased with the increase of concentration and soaking time however, laggards were not observed in T. aestivum. Clumping and chain formation of chromosomes at metaphase was also noticed in P. minor.