"PCR-karyotype" of human chromosomes in somatic cell hybrids

Amplification of human DNA sequences in 16 monochromosomal somatic cell hybrids containing different human chromosomes were performed by the polymerase chain reaction (PCR) using primer directed at human-specific regions of Alu or L1, the two major classes of interspersed repetitive sequences (IRS-PCR). A chromosome-specific pattern of amplification products was observed on agarose gels run with ethidium bromide, producing a "PCR-karyotype." This simple gel analysis provides a rapid method for identifying and monitoring the human chromosomal content of monochromosomal somatic cell hybrids without conventional cytogenetic analysis. Hybrids containing multiple human chromosome produce complex gel patterns, but identification of chromosome content can be achieved by hybridization of PCR products against a reference panel of monochromosomal or highly reduced hybrids representing each human chromosome. This dot-blot method also enables identification of human marker chromosomes or translocated pieces in hybrids that are not identifiable by cytogenetic methods. These IRS-PCR methods should greatly reduce the need for more laborious cytogenetic, isozyme, and Southern blot characterizations of human-rodent cell hybrids.     

Responses of transgenic maize hybrids to variant western corn rootworm larval injury

Abstract:  In 2005 and 2006, transgenic insecticidal maize hybrids (YieldGard Rootworm, MON 863, Cry3Bb1, Vector ZMIR 13L) were evaluated for their ability to limit root injury caused by western corn rootworm ( Diabrotica virgifera virgifera LeConte) larval feeding. Hybrids in each year of the experiment were planted in plots that had been devoted to a trap crop (late-planted maize interplanted with pumpkins) the previous growing season. All maize hybrids were provided by Monsanto Company and the genetic backgrounds remain unknown to the investigators. In 2005, the experiment was conducted in Urbana, Illinois. Urbana is located in east central Illinois, an area of the state in which a variant of the western corn rootworm has overcome the pest management benefits of crop rotation. Variation in root injury was noted across the maize hybrids in 2005 and the level of pruning increased from 20 July to 9 August for most hybrids. In 2006, the experiment was conducted in two locations, Monmouth and Urbana, Illinois. Monmouth is located in north-western Illinois and is within an area of the state in which densities of the variant of the western corn rootworm are lower than in east-central Illinois. In 2006, variation in root protection was again observed across the maize hybrids. Root injury differences among the hybrids were more prominent at the Urbana site. Similar to the previous year, root injury increased from the third week in July to the first week of August at both locations with this increase most noticeable at the Urbana location. We hypothesize that the variant western corn rootworm may be able to inflict more root injury to these transgenic insecticidal maize hybrids than the non-variant population of this species.     

Cytological and molecular characterization of oat x maize partial hybrids

In cereals, interspecific and intergeneric hybridizations (wide crosses) which yield karyotypically stable hybrid plants have been used as starting points to widen the genetic base of a crop and to construct stocks for genetic analysis. Also, uniparental genome elimination in karyotypically unstable hybrids has been utilized for cereal haploid production. We have crossed hexaploid oat (2n=6x=42, Avena sativa L.) and maize (2n=2x=20, Zea mays L.) and recovered 90 progenies through embryo rescue. Fifty-two plants (58%) produced from oatxmaize hybridization were oat haploids (2n=3x=21) following maize chromosome elimination. Twenty-eight plants (31%) were found to be stable partial hybrids with 1–4 maize chromosomes in addition to a haploid set of 21 oat chromosomes (2n=21+1 to 2n=21+4). Ten of the ninety plants produced were found to be apparent chromosomal chimeras, where some tissues in a given plant contained maize chromosomes while other tissues did not, or else different tissues contained a different number of maize chromosomes. DNA restriction fragment length polymorphisms (RFLPs) were used to identify the maize chromosome(s) present in the various oat-maize progenies. Maize chromosomes 2, 3, 4, 5, 6, 7, 8, and 9 were detected in partial hybrids and chromosomal chimeras. Maize chromosomes 1 and 10 were not detected in the plants analyzed to-date. Furthermore, partial self-fertility, which is common in oat haploids, was also observed in some oat-maize hybrids. Upon selfing, partial hybrids with one or two maize chromosomes showed nearly complete transmission of the maize chromosome to give self-fertile maize-chromosome-addition oat plants. Fertile lines were recovered that contained an added maize chromosome or chromosome pair representing six of the ten maize chromosomes. Four independently derived disomic maize chromosome addition lines contained chromosome 4, one line carried chromosome 7, two lines had chromosome 9, one had chromosome 2, and one had chromosome 3. One maize chromosome-8 monosomic addition line was also identified. We also identified a double disomic addition line containing both maize chromosomes 4 and 7. This constitutes the first report of the production of karyotypically stable partial hybrids involving highly unrelated species from two subfamilies of the Gramineae (Pooideae — oat, and Panicoideae — maize) and the subsequent recovery of fertile oat-maize chromosome addition lines. These represent novel material for gene/ marker mapping, maize chromosome manipulation, the study of maize gene expression in oat, and the transfer of maize DNA, genes, or active transposons to oat.Joint contribution of the Minnesota Agricultural Experiment Station and USDA-ARS. Scientific journal series paper No. 21 859 of the Minnesota Agricultural Experiment Station. Mention of a trademark or proprietary product does not constitute a guarantee or warranty by the USDA-ARS or the University of Minnesota and does not imply approval over other products that also may be suitable     

Transgenic maize hybrids as a tool to control Sesamia cretica Led. compared by conventional method of control on normal hybrids

This study was conducted on Bt and normal maize hybrids during two successive sowing dates, early and late summer plantation. The Sesamia cretica larvae were observed for the first time on first week of April on different commercial maize hybrids. While there was no larva/plant on transgenic maize hybrids, YieldGard and Ageeb-YG, while larvae were found on last week of July by a very little number on maize hybrids for late one. In Early summer planting date showed highly significant differences in reduction percent, between tested compounds and control one, in the pink corn borer population. The bio-residual activity of the NeemAzal-T/S 0.5% on TWC310 was significantly less pronounced, i.e. 75.5%, than other tested ones; the highest percentage of reduction, 86.6%, was achieved by Chlorophan 0.005% on SC2031, followed by NeemAzal-T/S 0.5% on SC2031 83.5% and Chlorophan 0.005% on SC2030 83.4%, respectively. Also the average reduction percentage of S. cretica larvae differed significantly from 69.6%, on NeemAzal-T/S SC2031, to 83.0%, on Chlorophan SC2031, in the late summer planting. The different treatments had positive effects on maize yield in the two planting dates. In the first plantation date, the increased rate of control ranged from 1.04 to 1.24, when NeemAzal-T/S acted on TWC310 and transgenic IYG, respectively. And in the second plantation date, rate of control obtained was 1.06–1.30, when NeemAzal-T/S acted on SC 2031 and transgenic IYG, respectively. The tested compounds played an important role in controlling the pink corn borer insect, S. cretica, and this study suggests that Cry 1Ab-expressing corn hybrids would provide a great value as a component of corn IPM in Egypt.     

Effect of planting dates and maize hybrids on the infestation with sorghum shootfly,Atherigona soccata Rondani and its effect on the yield

Abstract

Twenty maize hybrids plus local were tested for their susceptibility to major pests of sorghum shootfly, Atherigona soccata Rondani under field conditions at Sohag governorate during three planting dates (20th April, 20th May and 5th July) sown during two tested years (2004 and 2005 seasons). The investigation showed that when the sowing dates were considered irrespective of the different years or the maize hybrids it was evident that during the first planting date (20th April), the maize hybrids harboured the highest levels of infestation with sorghum shootfly, Atherigona soccata Rondani. The second planting date (20th May) was the least affected while the third planting date (5th July) received moderate levels of infestation. Results also showed that the average infestation of Atherigona soccata during both seasons was a significantly negative correlation affected by the sowing dates and maize hybrids. The yield of maize hybrids negatively correlated with percentage of infestation by Atherigona soccata. The highest yield obtained from the maize hybrids was slightly infested by Atherigona soccata and sown during the planting date of 20th May. Therefore, the second sowing date (20th May) may be recommended as a proper cultivating date for maize hybrids as it brought about reduction not only in infestation but was also a good method to reduce chemical control. Our results indicated that none of the maize hybrids evaluated were found to be resistant to Atherigona soccata attack but maize hybrids were significantly different in their susceptibility and less than of a local control to the infestation with Atherigona soccata and divided into four groups, the first one was highly susceptible including Giza Baladi; the second group was susceptible including ten maize hybrids namely, Hf 155, Bionear 3062, Bionear 30k8, Nagah 18, Watania 4, HC 326, HC 327, Nafratity, Watania 1 and Bionear; the third group was low resistance including six maize hybrids namely, Hf 122, Hf 123, Hf 129, Hc314, Hc 325 and Hc 352; while the fourth group included Hf 10, Hf 124, Hc 311and Hc 324 and moderately resisted the Atherigona soccata infestation. These may serve as a good material for growing in the areas where the pest is a problem.     

Effects of nitrogen deficiency on photosynthetic traits of maize hybrids released in different years

BACKGROUND AND AIMS: New maize (Zea mays) hybrids outperformed old ones even at reduced N rates. Understanding the mechanisms of the differences in performance between newer and older hybrids under N deficiency could provide avenues for breeding maize cultivars with large yield under N deficiency, and reduce environmental pollution caused by N fertilizers. METHODS: N deficiency effects on grain weight, plant weight, harvest index, leaf area and photosynthetic traits were studied in the field for six maize hybrids released during the past 50 years to compare their tolerance and to explore their physiological mechanisms. KEY RESULTS: N deficiency decreased grain yield and plant weight in all hybrids, especially in the older hybrids. However, there was no significant difference in harvest index, rate of light-saturated photosynthesis (Psat) 20 d before flowering, leaf area or plant weight at flowering between the N-deficient and control plants of all hybrids. Dry matter production after flowering of the N-deficient plants was significantly lower than that of the control plants in all hybrids, especially in the older hybrids, and was mostly due to differences in the rate of decrease in photosynthetic capacity during this stage. The lower Psat of the older hybrids was not due to stomatal limitation, as there was no significant difference in stomatal conductance (gs) and intercellular CO2 concentration (Ci) between the hybrids. N deficiency accelerated senescence, i.e. decreased chlorophyll and soluble protein contents, after anthesis more for the earlier released hybrids than for the later ones. N deficiency decreased phosphoenolpyruvate carboxylase (PEPCase) activity significantly more in older hybrids than newer hybrids, and affected the maximal efficiency of PSII photochemistry (Fv/Fm) only in the old hybrids and at the late stage. CONCLUSIONS: Compared with older (earlier released) hybrids, newer (later released) hybrids maintained greater plant and grain weight under N deficiency because their photosynthetic capacity decreased more slowly after anthesis, associated with smaller non-stomatal limitations due to maintenance of PEPCase activity, and chlorophyll and soluble protein content.     

喷施脱水剂对不同熟期夏玉米脱水特性和籽粒品质的影响

目前,我国种植的夏玉米品种收获时籽粒含水率过高,限制了玉米机械粒收技术的发展。喷施脱水剂可以调控作物籽粒灌浆生理过程,降低收获时的籽粒含水率。本试验研究了喷施脱水剂对不同熟期夏玉米品种脱水过程、收获期籽粒含水率和籽粒品质的调控作用。结果表明: 喷施脱水剂减少了玉米各器官的干物质积累量,促进了植株向籽粒中的干物质转移,提高了收获指数,而且对籽粒品质没有显著影响。相关性分析显示,籽粒脱水速率与各器官脱水速率呈正相关,喷施脱水剂后籽粒脱水速率与茎鞘脱水速率呈极显著正相关。喷施脱水剂在产量没有显著降低的前提下提高了总脱水速率,缩短了开花期至生理成熟期的时间,增加了生理成熟期到收获的时间,有利于后期籽粒含水率的进一步降低,为玉米机械粒收提供了更大的可能性。不同熟期夏玉米品种喷施脱水剂进行机械粒收的经济效益与机械穗收相比没有显著差异,中晚熟品种的经济效益高于早熟品种。因此,收获前合理喷施脱水剂可以作为玉米机收籽粒的一种可行性配套技术。     

Aberrant microspore development in hybrids of maize x Tripsacum dactyloides.

Cytogenetic investigations of meiosis in hybrids between maize and Tripsacum have been well documented; however, the inherent problem of male and female sterility has not been addressed either on a genetic or cytogenetic level. The purpose of this cytological study was to identify some of the probable causes of male sterility in maize x Tripsacum dactyloides hybrids. Disturbances in pollen development of maize x T. dactyloides hybrids, derived from both diploid (2n) and tetraploid (4n) Tripsacum sources, were commonly observed. Anomalies in the development of the microspore apparently occurred because of a failure of the chromosomes to congregate at the metaphase plate, development of a tripolar spindle, and failure of cytokinesis at the first and second meiotic divisions. Phenotypic features of abnormal microspore development were the maturation of large pollen grains, "Siamese" pollen grains, the occurrence of variable invaginations, and a nuclear budding-type behavior. These abnormalities were not observed in the 56-chromosome amphidiploid or the 38-chromosome backcross generations.     

Decomposition dynamics and structural plant components of genetically modified <Emphasis Type="Italic">Bt</Emphasis> maize leaves do not differ from leaves of conventional hybrids

The cultivation of genetically modified Bt maize has raised environmental concerns, as large amounts of plant residues remain in the field and may negatively impact the soil ecosystem. In a field experiment, decomposition of leaf residues from three genetically modified (two expressing the Cry1Ab, one the Cry3Bb1 protein) and six non-transgenic hybrids (the three corresponding non-transformed near-isolines and three conventional hybrids) was investigated using litterbags. To elucidate the mechanisms that cause differences in plant decomposition, structural plant components (i.e., C:N ratio, lignin, cellulose, hemicellulose) were examined. Furthermore, Cry1Ab and Cry3Bb1 protein concentrations in maize leaf residues were measured from harvest to the next growing season. While leaf residue decomposition in transgenic and non-transgenic plants was similar, differences among conventional cultivars were evident. Similarly, plant components among conventional hybrids differed more than between transgenic and non-transgenic hybrids. Moreover, differences in senescent plant material collected directly from plants were larger than after exposure to soil for 5 months. While the concentration of Cry3Bb1 was higher in senescent maize leaves than that of Cry1Ab, degradation was faster, indicating that Cry3Bb1 has a shorter persistence in plant residues. As decomposition patterns of Bt-transgenic maize were shown to be well within the range of common conventional hybrids, there is no indication of ecologically relevant, adverse effects on the activity of the decomposer community.     

Hydraulic lift in drought-tolerant and -susceptible maize hybrids

Hydraulic lift was investigated in a greenhouse study involving two drought-tolerant maize (Zea mays L.) hybrids (TAES176 and P3223) and a drought-susceptible hybrid (P3225) during the flowering stage. Root systems were grown in two soil compartments – a drier upper soil and a wetter deep soil. The plants were shaded for 3 h during the daytime. Soil volumetric water content (Ø_v) in the upper pots was measured with time domain reflectometry (TDR) before and after shading. An increase in Ø_v in the upper pot was detected with TDR in the drought-tolerant hybrids following 3 h of shading, but not in the drought-susceptible hybrid. Furthermore, water exuded from roots in the top soil layers was greater in the more drought-tolerant TAES176 than in P3223 (489 vs. 288 g per pot in 3 h, P<0.005). The sizable amount of water from hydraulic lift allowed TAES176 to reach a peak transpiration rate 27–42% higher than the drought-susceptible hybrid P3225 on the days when the evaporative demand was high. To our knowledge, this is the first experiment that reveals a significant surge of transpiration due to hydraulic lift following midday shading. Hydraulic lift also prevented soil moisture depletion in the upper pots with TAES176, but not with P3223 or P3225. Root characteristics may be responsible for differences in hydraulic lift of the three maize hybrids. There were 2.3–3.3-fold more primary roots in the deep moist soil in P3223 and TAES176 than in P3225 that may enable these hybrids to absorb and transport water at faster rates. Therefore, more water can be exuded into the upper drier soil when transpiration is suppressed by shading. Larger primary roots (20–28% larger diameter) and a higher root volume in the upper soil in TAES176 and P3223 than in P3225 may contribute to higher root hydraulic conductance and greater water efflux from the roots. The negligible hydraulic lift in P3225 may also relate to higher night-time transpiration of the hybrid. This report has documented, for the first time, the existence of genetic variations in hydraulic lift among maize hybrids and links between hydraulic lift and drought tolerance within maize plants. It appears that one of drought tolerance mechanisms in maize may lie in the extent of hydraulic lift.     

Transmission of maize chromosome 9 rearrangements in oat-maize radiation hybrids.

Oat-maize radiation hybrids are oat (Avena sativa L.) plants carrying radiation-induced subchromosome fragments of a given maize (Zea mays L.) chromosome. Since first-generation radiation hybrids contain various maize chromosome rearrangements in a hemizygous condition, variation might be expected in the transmission of these rearrangements to subsequent generations. The transmission and integrity of maize chromosome 9 rearrangements were evaluated in progenies of 30 oat-maize radiation hybrids by using a series of DNA-based markers and by genomic in situ hybridization. Maize chromosome 9 rearrangements were reisolated by self-fertilization in 24 of the 30 radiation hybrid lineages. Normal and deleted versions of maize chromosome 9 were transmitted at similar frequencies of 9.1% and 7.6%, respectively, while intergenomic translocations were transmitted at a significantly higher frequency of 47.6%. Most lines (93%) that inherited a rearrangement had it in the hemizygous condition. Lines with a rearrangement in the homozygous state (7%) were only identified in lineages with intergenomic translocations. Homozygous lines are more desirable from the perspective of stock maintenance, since they may stably transmit a given rearrangement to a subsequent generation. However, their isolation is not strictly required, since hemizygous lines can also be used for genome mapping studies.     

Prediction of maize double-cross hybrids using the best linear unbiased prediction with microsatellite marker information

We examined the usefulness of the best linear unbiased prediction associated with molecular markers for prediction of untested maize double-cross hybrids. Ten single-cross hybrids from different commercial backgrounds were crossed using a complete diallel design. These 10 single-cross hybrids were genotyped with 20 microsatellite markers. The best linear unbiased prediction associated with microsatellite information gave relatively good prediction ability of the double-cross hybrid performance, with correlations between observed phenotypic values and genotypic prediction values varying from 0.27 to 0.54. Taking into account the predictions of specific combing ability, the correlation between observed and predicted specific combining ability varied from 0.50 to 0.88. Based on these results, we infer that it is feasible to predict maize double-cross hybrids with different unbalance degrees without including any prior information about parental inbreed lines or single-cross hybrid performance.     

Detection of the apomictic mode of reproduction in maize-Tripsacum hybrids using maize RFLP markers

Polyploid plants in the genus Tripsacum, a wild relative of maize, reproduce through gametophytic apomixis of the diplosporous type, an asexual mode of reproduction through seed. Moving gene(s) responsible for the apomictic trait into crop plants would open new areas in plant breeding and agriculture. Efforts to transfer apomixis from Tripsacum into maize at CIMMYT resulted in numerou intergeneric F₁ hybrids obtained from various Tripsacum species. A bulk-segregant analysis was carried out to identify molecular markers linked to diplospory in T. dactyloides. This was possible because of numerous genome similarities among related species in the Andropogoneae. On the basis of maize RFLP probes, three restriction fragments co-segregating with diplospory were identified in one maize-Tripsacum dactyloides F₁ population that segregated 1∶1 for the mode of reproduction. The markers were also found to be linked in the maize RFLP map, on the distal end of the long arm of chromosome 6. These results support a simple inheritance of diplospory in Tripsacum. Manipulation of the mode of reproduction in maize-Tripsacum backcross generations, and implications for the transfer of apomixis into maize, are discussed.     

Biodetrimental effects of the Corn hybrids,Neemazal-T/S and Chlorphan 48% on the pink corn borer Sesamia cretica Led.

The aim of this study was to investigate influence of transgenic and commercial corn hybrids on the behaviour and feeding activity of the pink corn borer (Sesamia cretica Led.). Food consumption was different according to feeding period and hybrids. The feeding ratio of S. cretica on maize hybrids was significantly different between transgenic and commercial hybrids. It appears from the results of this experiment that the antifeeeding activity of transgenic hybrids had greater effect than commercial hybrids one. Thus, it is apparent from these results that transgenic maize was unsuitable because larvae were dead, but the other commercial hybrids were preferable. It can be concluded from the data that the feeding on different maize hybrids had different effects on certain biological aspects of pink corn borer. The impact of untreated Bt. Corn; 0.5% water emulsion of Neem-Azal-T/S and 0.005% water emulsion of Chlorpyrifos insecticides on the behaviour and feeding activity of the pink corn borer S. cretica has been studied. The data showed that larvae stopped feeding from the time when it was fed on untreated Bt. corn or/and non Bt. Corn treated with NeemAzal-T/S 0.5%. The results indicated that Bt. corn, NeemAzal-T/S 0.5% and Chlorphan 48% (0.005%) possessed the toxic effect on pink corn borer S. cretica. According to results, it could be stated that the tested compounds can play an important role in controlling the pink corn borer.     

The sugar,biomass and biofuel potential of temperate by tropical maize hybrids

The demand for biofuels has created a market for feedstocks to meet future energy requirements. Temperate × tropical maize (Zea mays L.) hybrids, which combine high biomass and fermentable stalk sugars, have yet to be considered as a biomass feedstock. Our objective was to evaluate biological potential, genetic variability and impact of nitrogen (N) on biomass, stalk sugar, and biofuel potential of temperate × tropical maize (TTM) hybrids. Twelve TTM hybrids, two grain and silage hybrids were grown in 2008, followed in 2009 by seven earshoot‐bagged TTM hybrids. In both years, they were grown with and without supplemental N (202 kg ha^?1) in Champaign, IL. Plants were sampled for total and partitioned biomass, and analyzed for concentration and content of sugar. The TTM hybrids were 40% taller, exhibited later reproductive maturity, greater flowering asynchrony, and remained green longer. All hybrids responded to supplemental N by producing more biomass and grain, a lower percent of biomass partitioned to stalk and leaf, whereas TTM also had a decreased concentration of sugar. Total average biomass yields were 24 Mg ha^?1 for both the TTM and grain hybrids. However, TTM partitioned 50% more biomass to the stalk and produced 50% more sugar, and had less than half the grain of the commercial hybrids, indicating grain production and sugar accumulation are inversely related. When grain formation was prevented by earshoot bagging, TTM hybrids produced, without supplemental N fertilizer, an average of 4024 kg ha^?1 of sugar, which was three‐ to four‐fold greater than the non earshoot‐bagged TTM and ear removed hybrid. Calculated estimates for ethanol production, considering the potential from sugar, stover and grain, indicate TTM can yield nearly the amount of ethanol per hectare as modern grain hybrids, but with a decreased requirement for supplemental fertilizer N.     

A more accurate method of comparing first-generation maize hybrids with their parents

Ohne Zusammenfassung

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A more accurate method of comparing first-generation maize hybrids with their parents

     

弱光胁迫对不同耐荫型玉米果穗发育及内源激素含量的影响

2010—2011年以耐荫性较弱的玉米杂交种豫玉22和耐荫性较强的玉米杂交种郑单958为材料,在抽雄前3 d开始进行弱光胁迫处理,吐丝后10 d恢复自然光照,研究弱光胁迫及光恢复对不同耐荫型玉米果穗生长发育及其内源激素含量的影响。结果表明:弱光胁迫下,玉米果穗生长发育明显减缓,穗长、穗粗和果穗干物质积累显著减小,秃尖度变大;穗行数、穗粒数和籽粒库容显著降低;吐丝期果穗顶部小穗子房发育停滞,已有败育迹象的籽粒在恢复自然光照后无明显改善;豫玉22果穗和籽粒性状在处理间的差异程度均大于郑单958。弱光胁迫下,两玉米杂交种果穗的ABA和ZR含量均升高,而GA含量和GA/ABA比值均降低;IAA含量和IAA/ABA比值在郑单958果穗中表现为升高,而豫玉22则表现为下降。     

Photosynthetic rate and yield formation in different maize hybrids

The relationship between photosynthetic rate and yield formation processes of the newer and older maize hybrids were investigated. Leaf area at flowering (source) and kernel number (sink) of the newer hybrids were greater than the older ones although their light-saturated photosynthetic rate (P_sat) were not greater than the older ones before flowering. After flowering, P_sat and chlorophyll content of the newer hybrids declined more slowly than the older ones. They not only distributed almost all photosynthates produced after flowering to grain but also reallocated some reserved photosynthates produced before flowering to grain. The newer hybrids exhibited greater grain mass than the older ones mostly because they could optimally regulate the photosynthetic rate and yield formation processes to maximize grain mass.