Agronomic and morphological characterization of Agrobacterium-transformed Bt rice plants

The agronomic and morphological characteristics of Agrobacterium-transformed rice plants carrying the synthetic cry1Ab or cry1Ac gene were investigated. Tremendous variations in plant height, seed fertility, grain size and other traits were seen in 80 T₁ lines, derived from 80 T₀ plants of 9 rice varieties. On average, about 33% T₁ lines had either morphological or agronomic variant plants. Most of the variations in T₁ plants had no significant correlation with transgene insertion and were proved heritable to their progenies. Genetic analysis in T₃ or T₄ generations showed some simple mutations such as chlorophyll deficiency and stunted plants were independent of transgene insertion and seemed to be controlled by a pair of single genes. However, in two independent transgenic progenies of Xiushui 11, all plants homozygous for transgenes showed dwarfism while all hemizygous and null segregants had normal plant heights. Two advanced homozygous Bt lines, KMD1 and KMD2, were developed from these two progenies. Comparison of the agronomic traits of KMD1 and KMD2 with their parent displayed marked differences among them in terms of seedling growth, tillering ability, yield components and yield potential. The genetic variation observed was generally not linked to the transgene locus and was ascribed to somaclonal variation, but other causes might also exist in particular cases. The results are discussed in the context of choosing appropriate transformation methodology for rice breeding programs. This revised version was published online in August 2006 with corrections to the Cover Date.     

Breeding for yield and quality in durum wheat

Three populations of an intervarietal durum wheat cross IWP5308/PDW208, F₅, F₅BIP₁ (population derived after intermating in F₂) and F₅BIP₂ (population derived after intermating in BIPF₁), were evaluated under three different agronomic environments for mean performance and stability of genotypes for grain yield, yield components and protein content. Though the biparental progenies indicated a higher mean performance, they did not differ significantly from progenies of the pedigree method for almost all characters. The biparental progenies, however, produced a higher number of stable genotypes for grain yield per plant, grains per ear and protein content. The F₅ population had a higher number of stable genotypes for 1000 grain weight and number of tillers per plant. The BIP progenies also had a higher number of genotypes with above average mean performance, and many were significantly higher than the checks WH896 and WH542, compared with F₅ progenies. Hence, in spite of high G x E interactions, the use of cycles of biparental mating and selection of top yielding lines on the basis of yield components can enable selection of stable genotypes with high protein content. Number of tillers per plant and 1000 grain weight were the yield component characters which made maximum contribution to phenotypic stability of the genotypes. This revised version was published online in August 2006 with corrections to the Cover Date.     

High regeneration potential in vitro of sunflower (Helianthus annuus L.) lines derived from interspecific hybridization

Successful selection of interspecific hybrid progenies with superior ability to regenerate shoots from apical meristems was performed in sunflower which now allows for the development of lines for improved biotechnological applications. Early generations of interspecific hybrids originating from crosses between the two H. annuus CMS lines ‘HA89’ and ‘Baso’, and 9 wild species were screened for their ability to regenerate in vitro. Evaluation of 36 progenies allowed to identify seven progenies from crosses involving H. mollis, H. giganteus, H. strumosus, and H. decapetalus which showed a significantly higher regeneration potential than the commercial hybrid ‘Albena’ regarding the number of shoots per explant. Among these progenies, 47.2 to 62.4% of explants produced shoots with an average of 2.3 to 3.5 shoots per cultured explant. Regeneration in vitro was significantly determined by the genotype. More than half of the investigated interspecific hybrids performed better than the inbred ‘HA89’ demonstrating that the high regeneration potential available in the wild species can be efficiently transferred to cultivated sunflower. The seven progenies with high regeneration potential in vitro were characterised by agronomic performance in the field. Two of the interspecific hybrids derived from H. strumosus and H. decapetalus not only showed a superior regeneration potential but also proved to be competitive to commercial hybrids with regard to important agronomic traits, e.g. fat content and TGW. This revised version was published online in July 2006 with corrections to the Cover Date.     

高丹草重组自交系群体的遗传变异与高产种质的创新

高丹草是一年生禾本科新型饲用牧草,提高其饲草产量是高丹草育种的重要目标.以高粱314A×棕壳苏丹草的F2:3遗传作图群体的后代材料建立的重组自交系(Recombinant inbred line,RIL)群体为材料, 对后代进行遗传变异和DNA多样性分析.结果表明:通过采用两种方法(性状和分子标记)进行同时选择,获得了9个高产的超亲重组后代.通过SSR标记分析RIL群体的DNA多样性,可知家系间存在较大的遗传变异,杂交重组产生的超亲高产种质分布在不同类群中.鉴定获得了2组SSR遗传距离接近0而且在主要农艺性状和产量等方面相似,只在抗性方面有差异的不同家系.农艺性状和抗性等鉴定试验筛选了4份性状优良的高丹草新种质并已有2份进入区域试验.     

Linkage between tolerance to high soil acidity and genetically high protein content in the kernel of wheat,Triticum aestivum L. and its possible use in breeding

Summary The winter wheat variety Atlas 66 has both the characteristics high protein content in the kernel and tolerance to high soil acidity. In a group of 121 F₄ lines, all derived from crosses with Atlas 66, a correlation coefficient of 0.39 was found between the two traits. A comparable group of 49 advanced generation lines, reselected for high protein content behaved similar, with a correlation coefficient of 0.37.The frequency distribution over the different categories of tolerance to soil acidity was tested within F₃ and F₄ populations of each two crosses between an Atlas 66-derived high-protein selection and a HRWW-variety. Both crosses slightly differed in frequency distribution but there were no differences between F₃ and F₄ distribution curves of one cross combination.Samples of varieties were tested, each sample having a phenotypically different protein content. The results indicated that protein content per se did not influence the level of tolerance to soil acidity. So it is concluded that although the two characteristics are genetically different they are linked to a certain extent.Individual regression lines were calculated for separate groups of F₄ selections derived from one cross combination. Two groups deviated from the majority and general pattern in that they showed a negative correlation between high protein and better tolerance. One of them differed significantly. A hypothesis is suggested to explain these exceptions.It is possible to screen lines for tolerance to high soil acidity in order to select within segregating populations for high protein content in the kernel, provided that positive selection is applied and that one of the parents combines both characteristics.     

山菠菜胆碱单加氧酶基因对棉花的遗传转化和耐盐性表达

胆碱单加氧酶（CMO）是渗透保护剂甜菜碱生物合成的关键酶之一。以棉花（Gossypium hirsutium L.）泗棉3号的下胚轴切段为外植体，利用农杆菌介导法将克隆自山菠菜（Atriplex hortensis）的AhCMO基因导入其中，通过组织培养胚状体发生途径获得转基因再生植株。以0.5%卡那霉素对再生苗筛选后，PCR检测抗卡那霉素棉苗确认阳性转化株，Southern和Northern杂交结果进一步证实外源基因已导入棉花并得到表达。在温室盆栽条件下，于2片真叶期对转AhCMO基因棉花及其转化受体泗棉3号施加0.5%的NaCl胁迫，15 d后发现其光合作用和植株生长被显著抑制，非转基因棉花泗棉3号的株高、鲜重和光合速率分别降低了57.6%、65.6%和69.9%，而转AhCMO基因的棉株分别降低了37.3%、54.6% 和47.9%。转AhCMO棉花所受盐害程度显著小于非转基因的棉株，说明AhCMO基因的导入和表达提高了转基因棉花的耐盐性。     

Inheritance of salt tolerance in rice

Summary The genetic behavior of salt tolerance was studied in artificially salinized conditions at the International Rice Research Institute.Divergent selection, carried out at a salinity level where the ECe was 15.2 mmhos/cm at 25 C in F₃ lines from two crosses confirmed the effects of salt tolerance on F₄ progeny with realized heritability values of 0.39 and 0.62, respectively.In a cross between two tolerant cultivars there was clear over-dominance for tolerance, despite the high environmental fluctuation which resulted in a low genetic response as indicated by a low but significant repeatability of 0.20–0.25, and many progeny lines more tolerant than the parents were recovered. The superior tolerance of these progenies compared to the parents was confirmed subsequently at 3 different salt levels. In the same experiment a cross between tolerant and susceptible cultivars produced some progeny of comparable tolerance with tolerant sources.In a 6×6 diallel cross experiment with two tolerant, moderate, and susceptible varieties each, both general and specific combining ability were significant.The findings indicate the possibility of breeding rices more tolerant than existing tolerant cultivars through cumulative crosses of tolerant cultivars. Further improvement can be attained by crossing highly tolerant lines with donors of good agronomic traits and pest and disease resistance.     

Properties of maternal haploid maize plants and potential application to maize breeding

Summary Presented are the results of a two-year study of haploid maize plants in the field. The haploids were produced with the aid of inducer line ZMS. In total, 604 and 1030 haploids were obtained and studied in the first and second years, respectively. Tassels of haploid plants were found to be almost completley sterile. Fertility of ears was studied by pollinating them with the pollen from diploid inbred lines, the cross resulting in almost all of the haploid ears carrying kernels. On average 27.4 kernels per ear of haploid plant were obtained in the first year of study and 26.3 in the second. These gave rise to normal diploid plants. This property allows genotypes selected at the level of haploid plants to be involved in breeding process. Unusual plants were found among haploids, phenotypically resembling homozygous lines. It was assumed that the plants had resulted from spontaneous chromosome doubling in haploids. The results of comparative studies of progenies of unusual plants and inbred lines derived from the same synthetic population are presented.     

Assessing genetic variation of maize (Zea mays) root DNA density under contrasting water supply

Optimizing root systems is one crucial point in drought tolerance breeding of plants. Introducing root-related traits to breeding programmes is time-consuming and laborious. Most of the commonly used methods are not suitable to be applied in a larger amount of plants. Here we present a study applying a DNA-based root phenotyping method (root DNA density; RDD) for phenotyping the root system of maize. Twenty-one maize inbred lines were investigated in a rain-out shelter experiment and 19 maize inbred lines in a greenhouse experiment under well-watered and drought conditions. Beside other commonly used root traits, agronomic traits of the plants were recorded and compared to RDD. Within root traits, RDD showed high significant genotypic variation and the highest repeatabilities of up to 72.4%. In contrast to most agronomic traits, repeatabilities increased under drought conditions. Values showed also good correlations between rain-out shelter and greenhouse trial, indicating the potential of this method for obtaining comparable results across different environments.     

Androgenetic green plants from winter rye, Secale cereale L., of diverse origin

Twenty varieties and breeders’ lines of winter rye, Secale cereale L., including Finnish, Polish, Russian, Swedish and Danish germplasm, were tested for their anther culture ability. To establish a reference point, the initial culture method was based on published results from rye anther culture. Following results from methodological studies, a modified protocol was tested on the same 19 winter rye lines. The modifications to the protocol included growth of mother plants under controlled greenhouse conditions, prolonged cold pretreatment of spikes for 3 weeks at 4°C, excision of anthers with microspores in the late uninucleate/binucleate developmental stage and induction on liquid medium supplemented with a buoyancy increasing component, Ficoll. With the modified protocol, the total production of green androgenetic plants increased nearly fivefold; it reached an average of 2.75% with the best line yielding 12.5%, while the proportion of green to albino plants increased by 50% to 25.1%. Eighteen rye lines showed green plant regeneration ability. Several genetically different winter rye lines with relatively good anther culture ability, yielding ≤ 1/% green plants per 100 anthers, were identified for the use in further methodical studies and for researching doubled haploidy in rye breeding.     

Testing for Tolerance to Aphids in Indian Mustard, Brassica juncea (L.) Czern and Coss.

Brassica juncea plants were screened for aphid tolerance using three methods. Seven and 14 days old seedlings grown in wooden trays in a glasshouse were kept between infector trays with heavy aphid infestation. There were no surviving plants in the susceptible genotypes 50 days after sowing but survival rates ranging between 80 and 90 % were observed for selection 44. The progenies of the surviving plants were tested under natural infestation in the field. Plants were scored for leaves without aphids 40 days after sowing, healthy plants at harvest and seed yield in g/m² For most of these lines, the aphids’ survival, life span and fecundity (nymphs produced per adult) were recorded after introducing five newly-born nymphs on excised fifth leaves cultured in petri dishes. Selections showing greater tolerance to aphids than the check cultivars were identified in the progenies of a cross RLM-514 × T-6342. It was concluded that the excised leaf method can provide a reliable measure of the lines’ reaction to aphids.     

The production of inbred lines of brussels sprouts with dominant S-alleles

Summary The best sources of Brussels sprout inbred lines with both good agronomic characters and high self-incompatibility are likely to be cultivars of reasonably good agronomic type which have not been too intensively selected. Comparison of three cultivars of different agronomic quality showed that the cultivars of poor and moderate quality had about 55% of plants with a dominant S-allele, but the most highly selected cultivar had only 25% of such plants. A programme of S-allele screening is suggested which incorporates the minimum number of tests required to determine whether or not a particular plant has a dominant S-allele. A survey of S-alleles present in commercial F₁ hybrids showed that the frequency of dominant S-alleles was only 19% in hybrids released prior to the end of 1971, but was 50% in hybrids released since 1971.     

A male sterile line with dominant gene (Ms) in cabbage (Brassica oleracea var. capitata) and its utilization for hybrid seed production

A male sterile plant, 79-399-3, was identified from a spring cabbage line 79-399 in 1979. Light-microscopic studies with paraffin section indicated that meiotic division stopped at the tetrad stage. The ratio of male sterile plants to fertile plants in the progenies of test crosses was 1:1 in five experiments conducted in 1982 and 1991–1994. However, some male sterile plants were sensitive and developed a very low number of viable pollen grains. When sensitive male sterile plants were selfed, the progenies segregated into male sterile and fertile plants at a ratio of 3:1. The male sterile plants from selfing of sensitive male sterile plants were propagated by tissue culture and crossed with different inbred lines. From the progenies of the crosses, populations with 100% male sterile plants were observed. The results indicated that homozygous dominant male sterile (MsMs) plants segregated from the selfed progenies. Populations with 100% male sterile plants and stable male sterility were developed as male sterile lines. Several favorable combinations were also selected by crossing the male sterile lines with inbred lines with high combining ability for desirable horticultural characters. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparison of QTLs mapped in RILs and their test-cross progenies of tropical maize for insect resistance and agronomic traits

Quantitative trait loci (QTL) affecting resistance to south-western corn borer Diatraea grandiosella (SWCB) and sugarcane borer Diatraea saccharalis (SCB) have been identified previously in F_2:3 lines and recombinant inbred lines (RILs) of tropical maize using restriction fragment length polymorphism (RFLP) analyses. Our objective was to determine whether QTLs identified in these generations are also expressed in test crosses (TC) of RILs. A population of 166 TC progenies was developed by crossing RILs from the cross CML131 (susceptible) × CML67 (resistant) with the unrelated, susceptible tester line CML216. Resistance to first-generation SWCB, measured as leaf-feeding damage (LFD) under artificial infestation, and other agronomic traits were evaluated in two environments for the TC progenies and three environments for 183 RILs. The correlation between line per se and TC performance was low for LFD and intermediate for most agronomic traits. Estimates of the genotypic variance and heritabilities were smaller in the TC progenies than in the RILs for all traits. Quantitative trait loci were identified using an RFLP linkage map with 136 loci. For LFD, four QTLs were detected in the TC progenies, of which two were in common with nine QTLs previously mapped in the RILs. Few QTLs for agronomic traits were common to the two types of progeny, because of the low consistency of QTL positions for all traits in RIL and TC progenies, the use of TC progenies should be considered in QTL mapping studies as the first step for marker-assisted selection in hybrid breeding.     

Salt-tolerance in Brassica juncea L. I. In vitro selection,agronomic evaluation and genetic stability

Summary In vitro selection of salt tolerant plants of Brassica juncea L. (Indian mustard) cv. Prakash has been accomplished by screening highly morphogenic cotyledon explant cultures on high NaCl media. Out of a total of 2,620 cotyledons cultured on high salt medium, 3 survived, showed sustained growth and regenerated shoots. They were multiplied by axillary bud culture on NaCl free medium. The salt-selected shoots retained salt tolerance following 3 month of growth and multiplication on control medium. While two of these somaclones flowered and set seeds, third one grew slowly, had abnormal leaf morphology and was sterile. The seed of the two fertile plants were sown in the field to raise R₁ segregating generation. Data were recorded for field, other agronomic components and oil content. The somaclonal lines, both selected salt-tolerant and non-selected, showed tremendous amount of variation for all the characters studied. One of the two tolerant somaclones invariably showed reduced height, longer reproductive phase and higher 1000 seed weight. Based on the agronomic performance of R₁ plants of these somaclones, some plants were selected and their progeny were evaluated for agronomic performance under standard field conditions and salt-tolerance in the greenhouse using sand pot culture method. Most of the lines bred true for their specific characteristics. In the greenhouse, selected salt-tolerant somaclones (SR-2 and SR-3) performed better for plant growth, yield and other agronomic traits at higher salt treatments, indicating thereby that salt-tolerance trait selected in vitro was expressed in the whole plants and is genetically stable and transmitted onto the progeny. The two tolerant lines, however, differed in their salt-tolerance during vegetative and reproductive phases as indicated by their salt-tolerance and stress susceptibility indices. The mechanism of salt-tolerance is not clear and needs to be further investigated.     

Breeding for resistance to cereal cyst nematode in wheat II. Use of test tube cultures in selection

Summary The use of soil. naturally infested with Heterodera avenae, to select resistant heterozygotes in backcross progenies of wheat, was tested for reliability. Selfed progenies from plants selected as resistant were cultured monoxenically in test tubes with nematodes hatched from single cysts, while backcross progenies from the same parent plants were grown in pots of naturally infested soil. Cyst counts were made after two months' growth. The results showed that over 50% of the backcross lines, screened in previous generations with naturally infested soil, had been erroneously selected as resistant. The test tube cultures clearly differentiated lines carrying resistance from those which were susceptible and corroborated results from pot tests.     

利用花药培养选育烤烟抗黑胫病突变体

首先在接种有不同浓度黑胫病菌发酵液活性组分的选择培养基上,获得高钾烤烟GK2的花药培养苗,并采用离体叶片法对这些花药培养苗进行黑胫病抗性鉴定,结果花药培养植株表现出不同程度的抗性;然后选其中6株抗性较高且农艺性状与GK2相近的单株,经秋水仙素加倍后收获种子,在黑胫病病圃及大田条件下于2016年对其R-1代、2017年对3个抗病性表现较好的R-2代植株进行黑胫病抗性及农艺性状测试,2年试验结果一致表明：在染色体加倍以后,所选植株的后代在基因型上已完全纯合,黑胫病抗性较GK2有较大提高且均可稳定遗传,是不同于原GK2的烤烟新品系,在生产上具有广泛应用前景。     

高密度对不同基因型夏玉米农艺特性、产量性状及耐密性的影响

为研究种植密度对不同夏玉米品种农艺性状、产量与耐密性的影响,在高密度（75000、90000株/hm²）条件下,对14个不同基因型夏玉米的农艺性状、产量构成因子以及耐密性系数等进行研究。结果表明,增密对14个不同基因型夏玉米品种生育期没有影响,株高、穗位高、茎粗、节间长度、茎粗系数、地上部干物质等农艺性状的差异达显著或极显著水平,穗位高系数、经济系数差异不显著,其中,株高平均增高1.23 cm,穗位高平均增高1.92 cm,第3节节间长度减小0.42 cm,‘NK718’、‘农大372’的地上部干物质质量增加,其余品种地上部干物质质量减小,减少幅度4.72%~23.99%,经济系数增加4.71%。75000 株/hm²条件下的产量、穗部性状与90000株/hm²密度条件相比,差异达极显著水平,在90000株/hm²密度条件下产量增加2.39%、穗粒数减少5.68%、百粒重降低1.17%。14个夏玉米品种相比,‘泽玉8911’、‘新单61’及‘豫单9953’耐密性系数值较大,适宜密植。     

In vitro screening and field evaluation of tissue-culture-regenerated sorghum (Sorghum bicolor (L.) Moench) for soil stress tolerance

Summary Sorghum bicolor (L.) Moench is generally quite sensitive to salt and acid (high aluminium) soil stresses, but quite tolerant of drought stress. As with any stress phenomenon, intra-specific variability exists within the genus. In vitro cell selection and somaclonal variation offer an alternative to traditional breeding methodology for generating improved breeding lines for hybrid development. A field selection protocol was developed for the three soil stresses and inter-stress evaluations were conducted in an effort to find multiple, stress-tolerant genotypes. The acid soil-drought stress, super-tolerant selections were located by the R₇ generation when exposed to a combined aluminium-drought stress field environment and when the regeneration population (number of regenerated lines from one callus source) was maintained at 15,000 plants or higher. A variant frequency of 0.1 to 0.2% for stress tolerance and acceptable agronomic traits among the surviving somaclones, provided an adequate number of phenotypes with desirable agronomic characteristics and a high level of soil stress tolerance. Subsequent research verified that the stress-tolerant regenerants had superior acid soil and drought stress tolerance to that of the donor parents, that their yield capabilities under stress were superior to their parents, and that their stress tolerance attributes were transferred in hybrid combinations. In vitro selection was not effective in increasing the number of field stress survivors. In fact, superior germplasms were developed from non-stressed callus or salt-stressed callus. In vitro selection reduced regeneration frequency and subsequent survival of plants under field stress. In vitro-stressed regenerants should be subjected only to non-stressed environments to maintain population numbers for field selection and thereafter should be subjected to stress environments during later (R₅₊) generations. The optimal strategy for the exploitation of somaclonal variation may be through short-term cell culture (< 12 months) with no attempt at in vitro selection.     

Variation in sweet potato for tolerance to some physical and biological stresses

Summary Sweet potato cultivars, breeding lines and unselected seedling clones were grown under 9 stress conditions in field and greenhouse; heavy soil, space competition, competition with maize, shading, flooding, drought, acidity, salinity, and weevil infestation. High root yield was considered to be a sign of stress tolerance. Stresses were shown to reduce the growth of shoots and roots, except flooding, which increased fibrous root production. The percentge of plants showing high stress tolerance varied from 0.35 to 30.90 for the different stress situations. Tolerances to stresses tended to occur together, as shown by low but significant correlations, significant chi-squares for numbers of plants with multiple stresses, and by low but significant correlations between yield and multiple stresses. Root flooding, space and fertility competition, and soil acidity appear to be the stress factors most closely related to production in heavy soils. Yet the magnitude of the correlations suggest that other factors or random variation also affect yield, and thus the predictive value of greenhouse stress test is presently limited.