Evidence for the genetic control of lysine catabolism in maize endosperm

A split pollination was used to produce normal (Su su su O2 o2 o2) and high lysine double mutant sugary opaque-2 (su su su o2 o2 o2) endosperms on the same ear of sugary opaque-2 maize plants. Amino acids were determined in the vascular sap of the ear peduncle. Lysine content in the sap was compared with lysine stored in both normal and sugary opaque-2 endosperm during kernel filling. Lysine content in the ear peduncle sap could account for all lysine found in both endosperms. Preformed lysine is highly catabolized in the normal endosperm, but not in the high lysine sugary opaque-2 endosperm. The rate of lysine breakdown appears to be an important mechanism by which the high lysine mutant controls lysine level in maize endosperm.     

Near-normal chiasma formation and maintenance in a short distal translocated segment in maize

Study of successful crossover pairing and chiasma formation is informatively extended to a very short translocated segment. Contrary to previous suggestion it now seems likely that the extreme distal region of the long arm of maize chromosome 1 is not deficient in intrinsic capacity for the initiation of crossover pairing. In addition, chiasmata formed in this short region appear to be efficiently maintained.     

The coupling of crossing over and homologous synapsis in maize inversions

Because fresh initiations of synapsis must occur for homologous synapsis of internal heterozygously inverted chromosome segments, attention has been directed at homologous synapsis and crossing over in overlapping paracentric inversions in the long arm of chromosome 1 of maize. In an earlier study with a relatively short inversion (where double crossovers within the inversion were rare), a recombination nodule (RN) was generally found at pachytene in reverse paired (homologously synapsed) inverted regions. Crossover frequency within the inversion, which could be independently estimated from analysis of bridge and fragment frequency at anaphase I and II, closely corresponded to crossover frequency estimated from observed RN frequency in pachytene inversion loops. These findings were consistent with the interpretation that establishment of homologous synapsis in this case is generally coupled to crossing over. This coupling suggests that there is very early commitment to the form of resolution of recombination intermediates that results in reciprocal recombination events instead of conversion only or other noncrossover events. This study examines another, larger paracentric inversion in the long arm of chromosome 1 that completely overlaps the first inversion. It is sufficiently longer than the first inversion that double crossover events are found within it with substantial frequency and interference considerations are feasible. This study confers additional insight into the interrelationships of synapsis and crossing over and the probable sequence in which the various involved processes usually occur. It raises the strong possibility that crossovers can be initiated during the alignment phase that precedes synapsis.     

Multiple forms of starch branching enzyme of maize: Evidence for independent genetic control

Purification of starch branching enzymes from kernels of two nonlinked mutants of maize, $\text{sugary}$ and $\text{amylose-extender}$, showed the basis of the two mutations to be associated with branching enzymes I and IIb, respectively. Branching enzyme I from $\text{sugary}$ kernels purified as nonmutant branching enzyme I, but had an altered pattern of activity when amylose was used as a substrate. In addition to the typical fall in absorbance at high wavelengths (550–700 nm) of the amylose-iodine complex, branching of amylose by $\text{sugary}$ branching enzyme I caused an increase in absorbance at low wavelengths (400–550 nm). Branching enzyme IIb was undetected in extracts of $\text{amylose-extender}$ kernels, while branching enzymes I and IIa appeared unaltered. Low umprimed starch synthase activity was also observed in DEAE-cellulose fractions of $\text{amylose-extender}$ maize, but this activity was regenerated by the addition of any branching enzyme.     

The desynaptic mutant of maize as a combined defect of synaptonemal complex and chiasma maintenance.

The phenotype of the desynaptic (dy) mutant of maize in microsporocytes at meiotic prophase was compared with normal microsporocytes of a closely related strain and with microsporocytes of a maize inbred line (KYS) assumed to be normal. Strikingly more univalents and open arms of bivalents were found in the mutant cells than in normal cells at diakinesis, and where there was heterozygosity for a distal knob (heterochromatic region), separation was usually equational, indicating the occurrence of normal crossing-over followed by failure of chiasma maintenance in the mutant. Differences found in the mutant by electron microscopy were a statistically significant wider dimension of the synaptonemal complex central region and also less twisting of synapsed configurations at pachytene. It is suggested that these are side-effect symptoms of a defect in the synaptonemal complex (or associated substance), which is expressed later as sporadic loss of chiasma maintenance.     

The modification of crossing over in maize by extraneous chromosomal elements

Summary Five regions of the maize genome were tested for their response to endogenous factors influencing recombination. These included heterochromatic B chromosomes and abnormal chromosome 10 as well as the sex in which recombination occurred.The frequency of recombination in the proximal A ₂-Bt and Bt-Pr segments of chromosome 5 was increased in the presence of B chromosomes, with the male meiocytes showing a greater response than the female meiocytes. In addition, experiments involving 0, 1, 2 and 4 B's revealed a dosage effect of B chromosomes on crossing over in chromosome 5. Recombination in the proximal Wx-Gl ₁₅ interval of chromosome 9 was found to be slightly higher than normal in male flowers when two B chromosomes were present. This increase was accompanied by a decrease in the adjacent Sh-Wx segment. Crossing over in the distal C-Sh segment and in the C-Sh-Wx-Gl ₁₅ regions of female flowers was unaffected by B's.Comparisons of plants heterozygous for abnormal chromosome 10 (K10 k10) and homozygous for the standard chromosome 10 (k10 k10) showed that abnormal 10 greatly enhances crossing over in the A ₂-Bt and Bt-Pr segments of chromosome 5. In contrast to the finding with B's, the effect is greater in female than in male sporocytes. K10 showed no significant effect on recombination in the C-Sh-Wx-Gl ₁₅ region of chromosome 9 except in male sporocytes, where there was a slight increase in the Sh-Wx region of 0 B K10 k10 plants and a possible interaction with B chromosomes to raise the level of recombination between Wx and Gl ₁₅. The fact that the regions adjacent to the centromere of chromosome 9 show little or no response to the presence of K10 indicates that the proximal heterochromatin of this chromosome differs qualitatively from that of other maize chromosomes. This conclusion is supported by a comparison of the effects of B chromosomes, K10 and sex on crossing over in chromosomes 5 and 9.Dedicated to Dr. M. M. Rhoades on the occasion of his seventieth birthday.     

Evidence for independent genetic control of the multiple forms of maize endosperm branching enzymes and starch synthases

Soluble starch synthase and starch-branching enzymes in extracts from kernels of four maize genotypes were compared. Extracts from normal (nonmutant) maize were found to contain two starch synthases and three branching enzyme fractions. The different fractions could be distinguished by chromatographic properties and kinetic properties under various assay conditions. Kernels homozygous for the recessive amylose-extender (ae) allele were missing branching enzyme IIb. In addition, the citrate-stimulated activity of starch synthase I was reduced. This activity could be regenerated by the addition of branching enzyme to this fraction. No other starch synthase fractions were different from normal enzymes. Extracts from kernels homozygous for the recessive dull (du) allele were found to contain lower branching enzyme IIa and starch synthase II activities. Other fractions were not different from the normal enzymes. Analysis of extracts from kernels of the double mutant ae du indicated that the two mutants act independently. Branching enzyme IIb was absent and the citrate-stimulated reaction of starch synthase I was reduced but could be regenerated by the addition of branching enzyme (ae properties) and both branching enzyme IIa and starch synthase II were greatly reduced (du properties). Starch from ae and du endosperms contains higher amylose (66 and 42%, respectively) than normal endosperm (26%). In addition, the amylopectin fraction of ae starch is less highly branched than amylopectin from normal or du starch. The above observations suggest that the alterations of the starch may be accounted for by changes in the soluble synthase and branching enzyme fractions.     

A possible role for the synaptonemal complex in chiasma maintenance

It is suggested that in addition to whatever function the synaptonemal complex may serve with respect to crossing over, it may serve a previously unsuspected function, i.e. one which normally assures regular meiotic chromosome disjunction in most organisms. This is the provision of the sister chromatid cohesiveness which is probably required for chiasma maintenance until anaphase I and may also be required in the centromere regions for maintenance of dyad integrity until anaphase II. Attention is directed to suggestive correlations of occurrence of normal synapsis through pachytene (homologous or non-homologous) with instances of sister chromatid cohesiveness at later meiotic stages and conversely of lack of normal synapsis with failure of sister chromatid cohesiveness. Chromosome behavior is compared in trisomies and in material homozygous for several meiotic mutants.     

Evidence for unequal crossing over in the evolution of the neurofilament polypeptide H.

Two allelic forms of the rabbit neurofilament protein H, designated H1 and H2, differ by approximately 6% in their electrophoretic mobilities. We have used techniques of peptide and nucleic acid analysis to determine that this difference is located in the central portion of the COOH-terminal tail, a region of the H protein that can project from the filament, and may form cross-bridges. This region comprises tandem amino acid motifs containing the sequence KSP, in which the serine residues are sites of phosphorylation. The sequence of this repetitive region of H2 is 132 nucleotides (44 amino acids) shorter than H1 and differs significantly in the arrangement of the repeated motifs. The difference suggests that both forms have evolved from a common ancestor over the past several million years by the process of unequal crossing over and that this process figured importantly in generating the repetitive region. In addition, the results indicate that this region is functionally redundant and that the length and structure of the tail of rabbit H resemble H from other mammalian species more closely than was suggested by a previously reported cDNA, on account of a single nucleotide difference.     

Chromosome-wide control of meiotic crossing over in C. elegans

A central event in sexual reproduction is the reduction in chromosome number that occurs at the meiosis I division. Most eukaryotes rely on crossing over between homologs, and the resulting chiasmata, to direct meiosis I chromosome segregation, yet make very few crossovers per chromosome pair. This indicates that meiotic recombination must be tightly regulated to ensure that each chromosome pair enjoys the crossover necessary to ensure correct segregation. Here, we investigate control of meiotic crossing over in Caenorhabditis elegans, which averages only one crossover per chromosome pair per meiosis, by constructing genetic maps of end-to-end fusions of whole chromosomes. Fusion of chromosomes removes the requirement for a crossover in each component chromosome segment and thereby reveals a propensity to restrict the number of crossovers such that pairs of fusion chromosomes composed of two or even three whole chromosomes enjoy but a single crossover in the majority of meioses. This regulation can operate over physical distances encompassing half the genome. The meiotic behavior of heterozygous fusion chromosomes further suggests that continuous meiotic chromosome axes, or structures that depend on properly assembled axes, may be important for crossover regulation.     

Evidence for genetic control of nondisjunction in man.

Data on factors associated with the occurrence of Down syndrome in a highly inbred population were evaluated to investigate the presence of a genetic control of nondisjunction in man. In Kuwait, close consanguinity occurs in 40% of marriages. In its main obstetric hospital, 20 trisomic Down babies out of 11,614 singleton births were delivered over a 12-month period. Chi-square analyses indicate the occurrence of Down syndrome to be linked to two independent factors: consanquinity of parents and maternal age. The relative risk is approximately four times greater for closely related than for nonrelated parents (P less than .005); a possible explanation for this is the existence of a gene that induces mitotic nondisjunction in the homozygous fertilized ovum. An alternative explanation is the existence of an autosomal recessive gene which results in meiotic nondisjunction in the homozygous parents. Consanguinity is usually perpetuated in certain families, or sections of the population, and parents in highly inbred families have a higher probability to be homozygotes for that gene.     

The pattern of pairing that is effective for crossing over in complex B-A chromosome rearrangements in maize

Special stocks involving complex B and A doubly translocated and recombined chromosomes were utilized to study the frequency and distribution of chiasmata under constrained conditions. The studies allow comparisons of frequency of pairing effective for crossing over in segments of different length and location in chromosomes that are present in disomic and trisomic quantity. Results provide new evidence for independent initiation of effective pairing in intercalary chromosome regions and suggest sequential events in the establishment of effective pairing, some of which may depend upon synaptic extension or two-by-two prealignment. Pairing frequency may depend directly on segment length under potentially competitive conditions. Evidence was not found for heterogeneity of pairing capacity within the regions studied.     

The pattern of pairing that is effective for crossing over in complex B-A chromosome rearrangements in maize

The study of the mechanism of meiotic homolog pairing, approached by comparing chiasma frequencies in rearranged segments that differ in relative length and intrachromosomal location, is substantially extended here. For the first time, two kinds of evidence were found that centers specialized for alignment pairing may exist in maize chromosomes: (1) for two segments, higher than average crossover frequency per unit length was maintained when these were located in several different chromosomal positions with respect to centromere and telomere, and in fact apart from their own normal centromeres and telomeres. High crossover frequencies in these segments regardless of position are considered to reflect innate capacity for alignment pairing due to relatively strong pairing center content. (2) For a short rearranged segment, chiasma frequency was drastically reduced, and evidence suggests that all of the chiasmata found there depended upon juxtaposition made possible by the completion of the zip-up pairing process in the other arms of the translocation configuration. This short segment is thought to be essentially devoid of pairing center content. It seems possible that crossover frequency depression in short rearranged segments may usually not be due, as commonly supposed, to mechanical difficulties inherent in formation of contorted configurations, but rather to absence of pairing centers within them and the relative rarity (compared to the normal sequence situation) of enabling zip-up pairing. Evidence also indicates that pairing which leads to crossing over must frequently occur between internal translocated segments and their normal sequence counterparts in a way which cannot be dependent upon zipping-up of two-by-two pairing initiated at or near telomeres. Pairing centers in maize are probably numerous and widely dispersed, since coarse direct proportionality is found when chiasma frequency is compared for an array of segment lengths.     

Chromosome-specific control of chiasma formation

A desynaptic mutant of Hypochoeris radicata, 2n = 8, has been found in a population from France. The mutant is remarkable in that at metaphase-I over 90% of PMC's have a pair of univalents while two pairs are found in only 1% of cells. Only chromosome IV is affected by the desynapsis which is controlled by a single recessive gene. Bivalent chiasma frequency in cells with a pair of univalents is higher than in cells with complete bivalent formation indicating partial dependence of cell chiasma number on the availability of some factor within the anther. The F₂, produced by full-sib mating, has a raised frequency of univalents in the chromosomes other than pair IV. The higher level results from enforced sib-mating of an obligate outbreeder and is not related to the action of the major gene. It is suggested that chiasma formation is controlled both polygenically and by major genes which operate on a hierarchical system. Some genes affect the behaviour of the entire complement while the action of others is specific to individual chromosomes.     

The control of chiasma distribution in rye

The chiasma conditions have been extensively analysed in two contrasting rye genotypes, one showing a normal and regular pattern of chiasma distribution and the other showing a highly abnormal and asymmetrical pattern of chiasma distribution. The analyses show that the pattern of chiasma distribution in the abnormal genotype conforms to the statistical expectations of randomness, and this is interpreted as being due to a breakdown of the processes which govern the distribution of chiasmata in normal rye genotypes. On the basis of these findings it is proposed that two independent and fundamentally different control systems are involved in the maintenance of efficient chiasma conditions in rye. One of these controls simply gives competence for chiasma formation, the other control system is evidently concerned with regulating the distribution of chiasmata. Analysis of two trisomic genotypes reveals that the conclusions relating to large samples of bivalents are also applicable to particular identifiable chromosomes. The results of brceding tests involving the two contrasting genotypes show that the control of chiasma distribution in rye has a complex genetic basis.This investigation was started while the author was in receipt of an Agricultural Research Council studentship at the Agricultural Botany Department, University College of Wales, Aberystwyth.     

Genetic and environmental components of chiasma control

Variation in chiasma frequency within and between individuals has been investigated in Schistocerca gregaria and Stethophyma grossum. By taking sequential samples of the same testis in S. gregaria it has been demonstrated that there is considerable variation in chiasma frequency between times, within individuals, and that this variation is not evenly distributed between individuals of the same population. The response and recovery patterns after exposure to successive temperature treatments also indicate a differential sensitivity of individuals within the experimental population. — An analysis of interfollicular variation in chiasma frequency has revealed significant differences between follicles within individuals both in S. gregaria and a Spanish population of S. grossum. A comparable analysis on individuals of S. grossum carrying supernumerary segments shows that the presence of such segments increases the amount of variation between follicles within individuals and also between individuals within the population. — The distribution of cell chiasma frequencies in S. gregaria is normal whereas in Stethophyma cell frequencies approximate to a Poisson distribution. The structurally different supernumerary segments, present in Austrian and Spanish populations of S. grossum, both increase mean chiasma frequency and they modify the between cell variance in different ways — the former interchromosomally and the latter intrachromosomally. — The differences in chiasma frequency between follicles and between times within individuals and the differential reaction to heat shock reflect differences in the pattern of genotype-environment interaction. Similarly it can be argued that the presence of polymorphisms involving supernumerary material may play an equivalent role by increasing the between cell or between bivalent variance within individuals, a unique form of genotype-environment interaction at the endophenotypic level. — Such variation in genotype sensitivity to environmental modification has important adaptive value especially in organisms which are subject to recurrent, often random, changes in their environment.     

Identification and genetic control of starch-degrading enzymes in maize endosperm

Three starch-degrading enzymes from liquid endosperm of maize have been separated by means of horizontal acrylamide gel electrophoresis. The three enzymes are tentatively identified as -amylase (zone 1), -amylase (zone 2), and -glucan phosphorylase (zone 3). Electrophoretic variants of these enzymes were found among ten inbred strains examined. Results of genetic crosses with respect to zone 2 amylase show that it is controlled by a pair of alleles (Amy-2 ^A and Amy-2 ^B) acting without dominance. It further appears that Amy-2 and Ct (catalase) are linked with 5% recombination frequency.This work was supported by the U.S. Atomic Energy Commission, under contract No. AT(11-1)-1338.     

The pattern of pairing that is effective for crossing over in complex B-A chromosome rearrangements in maize II

The problem of meiotic homologue pairing is approached by comparing chiasma frequencies in rearranged chromosome segments that differ substantially in relative length and intrachromosomal location. Results are consistent with affirmative answers to some questions previously raised: (1) whether there may be an underlying direct relationship between frequency of pairing and length of segment, (2) whether pairing commonly can be initiated independently in intercalary regions, and (3) whether there also can be a role for extension of pairing in adjoining regions for the establishment of pairing in intercalary regions, which requires pairing partner change. In addition, results here suggest that there may be: (1) greater capacity for establishment of pairing of more distal compared to proximal regions in a way that may also be dependent on their lengths, at least when these are relatively short, and additionally in a way which cannot be attributed to special properties of telomeres, (2) nearly random distribution of pairing of any two genetically long intercalary region representatives where three are present, without regard to the matching of the remainder of the chromosomes involved, and (3) a strong tendency for change of pairing partner in long distal segments when these are present in triplicate. Although sharp heterogeneities of pairing capacity were not found, it is suggested that they may exist with spacing too close for easy detection with the resolving power available.     

Genetic and environmental components of chiasma control

A disruptive selection programme for high and low chiasma frequency over four generations has produced two lines of Schistocerca gregaria with mean values of 21.6 and 18.0, respectively. The mean of the base population was 20.4. Realised heritability estimates for both the high and low lines were 27.4% and 48.8% respectively; only the latter value is significant. The non-significant value in the high line has resulted either from unidirectional dominance or because the genes responsible for high chiasma values themselves undergo a higher recombination rate in high frequency individuals and thus reduce the response to selection. — It is suggested that the genes governing chiasma frequency can be regarded as mainly neutral due to the large additive genetic component, at least in low chiasma frequency types. Accordingly, chiasma frequency variation and its adaptive significance may not be such an important component of fitness as has been proposed in the past.