Surrogate transformation of perennial ryegrass, Lolium perenne, using genetically modified Acremonium endophyte.

Summary Conditions have been developed for transforming protoplasts of the perennial ryegrass endophyteAcremonium strain 187BB. Unlike most other ryegrass endophytes, this strain does not produce the lolitrem B neurotoxin and is therefore suitable as a host for surrogate introduction of foreign genes into grasses. Transformation frequencies of 700–800 transformants/g DNA were obtained for both linear and circular forms of pAN7-1, a hygromycin (hph) resistant plasmid. Up to 80% of the linear transformants were stable on further culturing but only 25% of the circular transformants retained hygromycin resistance. Integration of pAN7-1 into the genome was confirmed by Southern blotting and probing of genomic digests of transformant DNA. Both single and tandemly repeated copies of the plasmid were found in the genome and both the number and sites of integration varied among the transformants. At least 13 chromosomes were identified in 187BB using contour-clamped homogeneous electric field (CHEF) gel electrophoresis. Probing of Southern blots of these gels confirmed that pAN7-1 had integrated into different chromosomes. The -glucuronidase (GUS) gene,uidA, was also introduced into 187BB by co-transformation of pNOM-2 with pAN7-1. GUS activity was detected by growing the transformants on plates containing 5-bromo-4-chloro-3-indolyl -D-glucuronic acid and by enzyme assays of mycelial extracts. Severalhph- anduidA-containing transformants were reintroduced into ryegrass seedlings and expression of GUS visualized in vivo, demonstrating that 187BB can be used as a surrogate host to introduce foreign genes into perennial ryegrass. Molecular analysis of fungal isolates from the leaf sheath confirmed that the pattern of pAN7-1 and pNOM-2 hybridizing fragments was identical to that observed in the fungus used as inoculum.     

热休克诱导鳙鱼四倍体的研究

采用热休克法研究了诱导鳙鱼四倍体的可能性和处理条件。结果表明:1)恰好在卵子开始第一次卵裂前用39—42℃水浴处理1—3分钟,可获得较高频率的四倍体;2)开始处理的时间至关重要,提前或推迟处理导致几乎全部死亡或四倍体频率大大下阵;3)热休克组的孵化率显著下降,畸形率明显升高;4)含高频率四倍体的热休克组的细胞分裂和胚胎发育延迟。     

Chromosomal instability in Lathyrus sativus L.

Summary In Lathyrus sativus (2n=14), variety LSD-1 shows an instability of somatic chromosome number which can be observed in root tip and shoot tip mitoses. In this variety, approximately 54% of the seedlings showed intra-individual variation in chromosome number ranging from 2n=14–3. This variability in chromosome number was recorded in approximately 60% of the dividing cells. Two seedlings were triploid with 21 chromosomes. Variation in chromosome number in somatic cells within individual plants is possibly controlled by genetic factors, which result in spindle abnormalities, chromosome degradation and minute chromosomes. The variation in chromosome number is probably responsible for the pollen polymorphism noted in this particular strain. The possible mechanism of intra-individual variability and the occurrence of the phenomenon vis-a-vis its applications are discussed.     

Multiple chromosomal interchanges in pearl millet

Summary Intercrossing and irradiation were successfully used in pearl millet (Pennisetum typhoides) to develop multiple interchanges involving up to the total complement of all the chromosomes in one complex. In interchange heterozygotes showing 12 + 1 II and 14, 90.9 and 87.8 per cent of the cells, respectively, had chromosome configurations other than that of 12 and 14 chromosomes. In general, the frequency of such cells resulting from breakdown of the expected complex configuration increased with the increase in the number of translocated chromosomes in the complex. The higher the number of chromosomes involved in the interchange ring, the higher were the pollen and ovule sterility. The results indicated that meiotic instability, deficiency-duplication gametes, and unequal distribution of chromosomes account for increased sterility of multiple interchange heterozygotes. Even though interchanges in pearl millet predominantly show the alternate type of segregation, sterility seems to be the major barrier for the exploitation of the multiple interchange method for gamete selection and the establishment of homozygous lines in this plant species.     

Analysis of the rye chromosome constitution and the amount of telomeric heterochromatin in the widely and narrowly adapted hexaploid triticales

Summary Investigations were made on the rye chromosome constitution and on the presence of telomeric heterochromatin in rye chromosomes of the 26 most widely and 24 most narrowly adapted triticale strains. Among widely adapted lines, 22 (85%) had a complete rye genome and four triticales only had chromosomal R-D genome substitutions. Twenty-three (96%) of the 24 most narrowly adapted triticales had substitutions between the chromosomes of the R and D genomes. The most widely adapted triticales accumulated fewer modified rye chromosomes in comparison to narrowly adapted lines. They had from one to three rye chromosomes with heterochromatic deletions: 46% of widely adapted lines had two modified rye chromosomes; 34% had three modified rye chromosomes, and 19% had a single modified rye chromosome. In widely adapted strains, the 1R, 4R, 5R and 6R modified chromosomes were observed; they were present in 80%, 73%, 50% and 11% of the cases, respectively. The most narrowly adapted triticales had from two to four modified rye chromosomes: 58% of the strains had three modified rye chromosomes; 29% had four modified rye chromosomes and 12% had two modified rye chromosomes. The modified 4R and 5R chromosomes were present in all of these lines. The 1R (modified), 6R (modified) and 7R (modified) were found in 83%, 25% and 16%, respectively, of the narrowly adapted strains.Results support the previous observations (Pilch 1980b) that a wide adaptation of hexaploid triticales is associated with the presence of the full potential of rye genome, and that it is independent of the amount of telomeric heterochromatin possessed by rye chromosomes.     

Schistosoma mansoni: Electrophoretic characterization of strains selected for different levels of infectivity to snails

Individual adult Schistosoma mansoni from strains selected for high or low infectivity to specific strains of the snail intermediate host, Biomphalaria glabrata, were subjected to enzyme electrophoresis on starch gels. Fourteen enzyme systems were analyzed in an attempt to find electrophoretic markers associated with genes for infectivity to snails. The S. mansoni strains were selected from different isolates from Puerto Rico in several strains of B. glabrata. Of an estimated 18 loci, 3 were polymorphic and the remainder monomorphic. For 1 of the 3 polymorphic enzyme loci, lactate dehydrogenase (Ldh, EC 1.1.1.27), phenotype frequencies were correlated with infectivity to snails. In schistosome strains of low infectivity, frequencies of the Ldh-N phenotype ranged between 0.56 and 0.69, while in strains of high infectivity, Ldh-N frequencies were typically 0.91 to 1.00. Whether the correlation is accidental or due to some form of association, such as chromosomal linkage, between the locus responsible for variation in lactate dehydrogenase and a gene for infectivity to snails remains to be determined.     

Age- and density-dependent growth within populations of the ectoparasitic digenean Transversotrema patialense on the fish host

Mills C. A. 1980. Age- and density-dependent growth within populations of the ectoparasitic digenean Transversotrema patialense on the fish host. Internationaljournal for Parasitology, 10: 287–291. Growth of the adult ectoparasitic digenean Transversotrema patialense on the fish host Brachydanio rerio is shown to be age-dependent, ceasing 15–20 days post infection. The vitelline glands expand greatly in size after infection from 1.25 % of cercarial area to 20.7 % of that of the mature adult. There is an increase in the occurrence of reproductive abnormalities in old parasites but this alone fails to account for the decline in egg production found as populations of T. patialense age. Growth of adult T. patialense is density-dependent with reduced growth at high initial parasite densities per host.     

Absence of repetitive DNA sequences associated with sex chromosomes in natural populations of the Trinidad guppy (Poecilia reticulata)

The genus Poecilia has been widely studied as a model for the evolution of sex chromosomes. In the course of molecular studies on population genetic structure and sexual selection in the Trinidad guppy, we examined our preparations for male-linked, repetitive DNA polymorphisms. We have not obtained any evidence of male-specific polymorphisms, in contrast to an earlier study. Our results have significant implications for theories on the evolution of sex chromosomes.Correspondence to: F. Breden     

Parthenogenesis and developmental constraints

The absence of a paternal contribution in an unfertilized ovum presents two developmental constraints against the evolution of parthenogenesis. We discuss the constraint caused by the absence of a centrosome and the one caused by the missing set of chromosomes and how they have been broken in specific taxa. They are examples of only a few well‐underpinned examples of developmental constraints acting at macro‐evolutionary scales in animals. Breaking of the constraint of the missing chromosomes is the best understood and generally involves rare occasions of drastic changes of meiosis. These drastic changes can be best explained by having been induced, or at least facilitated, by sudden cytological events (e.g., repeated rounds of hybridization, endosymbiont infections, and contagious infections). Once the genetic and developmental machinery is in place for regular or obligate parthenogenesis, shifts to other types of parthenogenesis can apparently rather easily evolve, for example, from facultative to obligate parthenogenesis, or from pseudoarrhenotoky to haplodiploidy. We argue that the combination of the two developmental constraints forms a near‐absolute barrier against the gradual evolution from sporadic to obligate or regular facultative parthenogenesis, which can probably explain why the occurrence of the highly advantageous mode of regular facultative parthenogenesis is so rare and entirely absent in vertebrates.