Relationship Between Phytohormones and Male Sterility in Thermo-Photo-Sensitive Genic Male Sterile (TGMS) Wheat

Summary Fertility alternation normally occurs in a novel type of thermo-photo-sensitive genic male sterile (TGMS) wheat whose recessive genes in the nucleus are expressed as virtually complete sterility under lower temperature and shorter daylight conditions, but as normal fertility under higher temperature and longer daylight conditions during the developmental period from meiosis of pollen mother cells (PMCs) to the early mononucleate stage of microsporogenesis. In order to provide more information about the mechanism(s) of fertility alternation in TGMS wheat, the relationship between fertility alternation and levels of phytohormones was analyzed by endogenous hormone quantification and exogenous hormone treatments in the present research. Sown at an earlier date with a relatively low temperature and short daylight, TGMS wheat was male sterile phenotypically. Pollen abortion mainly occurred at the mononucleate to bi-nucleate stages of microsporogenesis and pollen abortion was mainly of the spherical type. Levels of endogenous indole-3-acetic acid (IAA), abscisic acid (ABA) and gibberellic acid (GA₃) in the anthers were at lower levels, and zeatin riboside (ZR) was higher. Sown at a later date, when the temperature during the critical period for fertility alternation was relatively high and the daylight was long, TGMS wheat was almost as fertile as the normal genotype and the contents of endogenous IAA, ABA and GA₃ in anthers were at higher levels, and the content of ZR was at a lower level. At the critical period for fertility alternation, treatments with exogenous IAA, ABA, GA₃ or aminoethoxy vinylglycine (AVG), an inhibitor of ethylene biosynthesis, decreased the frequency of sterile pollen grains and increased the frequency of self-pollinated fertile florets. Treatment with exogenous 2-chloroethyl phosphonic acid (CEPA), a releaser of ethylene, increased the frequency of sterile pollen grains and decreased the frequency of self-pollinated fertile florets. It was thus suggested that the fertility alternation of TGMS wheat was closely related to levels of phytohormones in the anthers, and changes of endogenous hormone levels were among the important factors responsible for the fertility alternation of TGMS wheat.     

Mutations blocking sensitivity to gibberellic acid promote ethylene-induced male sterility in wheat

Summary Ethylene is known to perturb normal reproductive development in wheat, particularly the development of functional pollen. Two experiments were carried out to test the hypothesis that increasing insensitivity to gibberellic acid (GA), conferred by the Rht genes, would be associated with increased male sterility in ethrel or Cerone^®-treated wheat. Wild type (WT=rht1/rht1, rht2/rht2), single dwarf (SD=Rht1/Rht1, rht2/rht2 or SD=rht1/rht1, Rht2/Rht2), and double dwarf (DD=Rht1/Rht1, Rht2/Rht2) near-isogenic lines in six genetic backgrounds were treated with ethrel or Cerone^® at the late tetrad to early uninucleate stage of pollen grain development. Ethrel induced pollen abortion in all genotypes but was highest for DD (41% above background) followed by SD (20%), and then WT genotypes (10%). Spikelet fertility decreased as the number of Rht alleles increased in response to ethrel or Cerone^® treatments. Expressed as a percent of controls, spikelet fertility was 56% for WT, 42% for SD, and 29% for DD. The consistent linear relationship between the number of Rht alleles and sensitivity of ethylene-induced male sterility suggests that GA and its recognition may exert a stabilizing effect in pollen development in the presence of stress or an ethylene shock.Paper No. 764 of the Department of Plant Breeding and Biometry, Cornell Agricultural Experiment Station.     

Hybrid seed production in safflower (Carthamus tinctorius) following the induction of male sterility by gibberellic acid

The objective of the present research was to develop an effective and practical method for hybrid seed production in safflower, by inducing male sterility with gibberellic acid (GA₃). Seeds of the spiny line ‘5‐154’ and the non‐spiny variety ‘Dincer 5‐118’ were sown in alternate rows and the plants of the non‐spiny female variety were exposed to various concentrations of GA₃ for male sterility induction. Three successive sprays (75, 82 and 89 days after sowing) of 100 p.p.m. of GA₃ to safflower buds of <0.5 cm diameter at a pre‐meiotic interphase stage resulted in reduced pollen viability from 81.6‐6.7% compared with the control. An average hybrid seed percentage of 80.7% (87.5% in the secondary heads), identified by the use of the spiny‐capitulum marker variety ‘5‐154′, was obtained. The GA₃ treatments did not significantly affect the production of viable achenes. The results showed that GA₃ could be successfully used as a chemical hybridizing agent in hybrid safflower production.     

赤霉素对盐胁迫抑制水稻种子萌发的缓解作用的蛋白质组分析

用粳稻日本晴(Oryza sativa L. cv.Nipponbare),研究了盐胁迫对水稻种子萌发的抑制作用和赤霉酸(GA₃)对盐胁迫的缓解作用;分别以H₂O (对照）,5 g L^-1 NaCl (处理I),5 g L^-1 NaCl + 100 μmol L^-1 GA₃(处理II)培养水稻种苗48 h,提取芽中的蛋白质,利用双向电泳(2-DE)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术分析了水稻蛋白质组的变化。结果表明,在盐胁迫条件下,日本晴种子的萌发显著受到抑制,而GA₃能显著缓解这种抑制作用;用ImageMaster软件分析2-DE凝胶,发现有4个蛋白质斑点表现出显著的变化,在盐胁迫下斑点S1、S2和S3表达下调而斑点S4消失,在GA₃与盐共处理时,这4个蛋白质点的表达均有不同程度的恢复;经MALDI-TOF MS分析,其中2个蛋白质斑点(S1,S3)分别被鉴定为isoflavone reductase-like蛋白与葡萄糖磷酸变位酶,这些蛋白可能与GA₃提高水稻耐盐性途径相关。     

The effect of gibberellic acid on fruit and seed set in crosses of garden and winter hardy Rosa accessions

Summary The effect of gibberellic acid (GA₃) was studied on a wide range of crosses between various garden roses and two hardy breeding lines. Although there were fewer seeds per fruit, fruit set was higher in most crosses when GA₃ was applied to the stigma at the rate of 250 ppm ten days after pollination. However, higher fruit set did not result in more seeds per pollination for many crosses and seemed to be related to the degree of male parentage. Both parents used had a major effect on pollination success. Recently developed Rosa cultivars have a narrow genetic base and the use of distantly related cultivars as parents would possibly give a high seed set and greater numbers of progeny.Contribution No. 335/90.08.01R, Experimental Farm, L'Assomption; Contribution No. Ro 44, Research Program Service, Ottawa.     

外源IAA、GA3和ABA影响不同穗型小麦分蘖发生的机制

以山农15和山农8355为材料,研究外源生长素(IAA)、赤霉素(GA₃)、脱落酸(ABA)对小麦分蘖发生的影响及其作用方式。结果显示,外源IAA和GA₃可抑制分蘖的发生,而外源ABA则仅减缓了分蘖发生速率。进一步分析外源激素对分蘖芽生长动态及分蘖节中内源激素变化的影响,发现外源IAA和GA₃完全抑制分蘖芽的生长,而外源ABA明显减缓了分蘖芽的生长速率。外源IAA提高分蘖节中IAA含量和IAA/ZT比,抑制ZT含量的提高;外源GA₃提高分蘖节中IAA含量、IAA/ZT比和ABA/ZT比,抑制ZT含量的提高;外源ABA提高分蘖节中ABA/ZT比,减缓了ZT含量的升高速率。相关性分析显示,分蘖芽的生长与ZT呈显著正相关,与IAA、IAA/ZT比和ABA/ZT比呈显著负相关,与GA₃和ABA呈不显著的负相关。试验结果表明,IAA和ZT在小麦分蘖发生过程中起关键作用,外源激素主要通过影响内源IAA、ZT含量以及IAA/ZT比和ABA/ZT比来影响分蘖芽的生长,进而调控小麦分蘖的发生。     

The Identification of a Gibberellic-Acid-Insensitive Gene in Secale cereale*

Among 16 dwarfing genes identified in wheat (Triticum aestiuvm L. em Thell.), four are known to be associated with insensitivity to the externally-supplied growth hormone gibberellin (GA). Rht1 and Rht2 (Reduced height 1 and 2, respectively) have been the most extensively used, because of their positive effect on yield. To increase the germplasm pool for dwarfism, a spring rye (Secale cereale) population (UC-90, CI-174) was selected because it contains high variability and any useful genes would benefit triticale and wheat as well. Seedlings of the CI-174 rye population were treated with 50 ppm of GA to identify any insensitive types. GA-insensitive and -sensitive seedlings were identified and, after three generations of selfing, GA-insensitive and -sensitive lines were fixed. Rye insensitive was crossed to a sensitive wheat and to rye and, reciprocally, insensitive wheat was crossed to sensitive rye. The results indicated that a GA-insensitivity dwarfism system similar to that originally found in wheat also operates in rye and appears to be under simple inheritance. Rye GA-insensitivity was expressed in triticale. Therefore, it is possible to transfer this new source of insensitivity and dwarfism into triticale and wheat.     

Chromosomal Location of Genes for Gibberellic Acid Insensitivity in 'Chinese Spring' Wheat by Tetrasomic Analysis

The tetrasomics of the homoeologous groups 2, 5 and 7 of‘Chinese Spring’wheat were, together with the euploid standard, screened at the seedling stage for sensitivity to exogenously applied gibberellic acid (GA³). Whilst the seedling length of lines tetrasomic for group 2 chromosomes were taller and those for chromosomes 5A, 5D and 7D shorter in both treatments (with and without GA³) compared to the euploid control, the remaining tetrasomics — 5B, 7A and 7B — were significantly shorter than the euploids in the GA variant only. These results suggest the presence of additional genetic factors for GA insensitivity on chromosomes of the groups 5 and 7 of hexaploid wheat. This corresponds with the localization of GA insensitive dwarfing genes on the homoeologous chromosomes 5R and 7R in diploid rye.     

Effect of light on endogenous levels of gibberellin and abscisic acid in seed germination of photoblastic weedy rice (Oryza sativa L.)

The effect of red (R) and R/far-red (FR) lights on endogenous gibberellin (GA) and abscisic acid (ABA) content was first investigated during the germination of photoblastic black-hulled weedy rice (PBWR) seeds. The R light-treated PBWR seeds germinated after 36–48 h and germination was increased to 63% at 72 h. However, the FR light-treated seeds after R light treatment, suppressed the R light effect showing only 11% germination even at 72 h after the light treatment. The PBWR seed treated with R light rapidly increased the endogenous level of GA₁ to about 200 times at 12 h before seed germination as compared with R/FR (control) which suppressed the effect of R light. The contents of other GAs like GA₁₂, GA₅₃, GA₁₉, GA₂₀, and GA₈ were not affected by the R light irradiation. These results showed that the major biosynthetic pathway of GAs in PBWR seeds is the early 13-hydroxylation pathway leading to GA₁, which was suggested to be physiologically active in the PBWR seed germination. The decrease in the level of ABA in the R light-treated seeds was greater than the R/FR light-treated seeds, indicating that the balance of endogenous GA1 and ABA is responsible for the induction of germination in the PBWR seed.     

Utilization of tom thumb source of pre-harvest sprouting tolerance in a wheat breeding programme

Summary Segegating populations of the cross Tordo (a spring white-grained Tom Thumb derivative) and Tr 454-16 (a spring white-grained Sonora-Gabo derivative) were studied for plant height, apparent -amylase synthesis (AAS) and response to gibberellic acid. Means, variances and heritabilities for these characters were computed and their association was studied by computing correlation coefficients. Plant height showed high heritability while AAS and response to GA₃ showed moderate heritability. Plant height was positively correlated with AAS and response to GA₃. The results pointed out the importance of using Tom Thumb as a source of pre-harvest sprouting tolerance in a wheat breeding programme, particularly in areas where wheat breeding is restricted to evolving white-grained cultivars.     

Factors responsible for levels of male sterility in photoperiod-sensitive cytoplasmic male sterile (PCMS) wheat lines

A `two-line system' using photoperiod-sensitivecytoplasmic male sterility (PCMS) caused by Aegilops crassa cytoplasm has been proposed as a newmeans of producing hybrid wheat. The PCMS line ismaintained by self-pollination under short-dayconditions (14.5 h light period), and F<sub>1</sub> seedscan be produced by outcrossing of the PCMS line witha pollinator under long-day conditions (15 h lightperiod). As the levels of male sterility in PCMSlines under the long-day conditions is a crucialfactor in determining hybrid purity of the F<sub>1</sub>seeds, a study was conducted into the effect ofseeding rate on male sterility in PCMS lines. Threedifferent density levels were tested using analloplasmic line of Japanese wheat cultivar `Norin 26'which exhibits PCMS. Levels of male sterility of thePCMS line increased at sparse planting, because tiller(ear) number per plant increased at low seedingdensity and late-appearing ears tended to exhibithigher levels of male sterility than early-appearingears. On the other hand, male sterility levels of thePCMS lines depended on genotype, e.g., the PCMS`Fujimikomugi' was completely male sterile, whereasthe PCMS `Norin 26' showed partial male sterility. APCMS line showing complete male sterility, such as thePCMS `Fujimikomugi', should produce F<sub>1</sub> seeds withhigh purity. However, the PCMS `Fujimikomugi' showeda lower female fertility. For practical use, it isnecessary to produce PCMS lines having high malesterility with high female fertility under long-dayconditions.     

An assessment of morphogenic and transformation efficiency in a range of varieties of potato (Solanum tuberosum L.)

Summary To provide a truly genotype-independent transformation system, it is necessary to be able to transform a wide range of potato genotypes. The ability to regenerate shoots in vitro was determined for 34 potato varieties using tuber disc explants. Following a culture regime used extensively in previous studies with the variety Desiree, half of the varieties could be regenerated from tuber discs and half could not. From a sample of varieties that could be regenerated from tuber discs, all but one variety gave transgenic plants. Twelve varieties were evaluated for the capacity to regenerate shoots from leaf and internode explants excised from in vitro grown plants. All of the varieties tested regenerated adventitious shoots. Leaf and internode explants from 5 varieties were subsequently used for transformation, and transgenic plants were produced from two potato varieties that did not give transgenic plants from tuber disc explants. Some varieties could not be transformed by either method, and will require modification of the in vitro regeneration and transformation system to be successful.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid     

Classification of GA response,Rht genes and culm length in Japanese varieties and landraces of wheat

Summary The GA response, Rht genes and culm length of 133 Norin varieties, 6 breeding lines and 16 landraces of Japanese wheat were investigated. Out of 133 Norin varieties tested, 103 were GA-insensitive and 30 GA-responsive. The 6 breeding lines were all GA-insensitive. Out of 16 landraces tested, 10 were GA-insensitive and 6 GA-responsive. Among the 10 GA-insensitive landraces, only Daruma had a Rht1 genotype. The other 9 had a Rht2 genotype. None of the landraces tested carried both Rht1 and Rht2 or Rht3. Out of the 103 GA-insensitive Norin varieties, 22 carried only Rht1, another 79 carried only Rht2, and only Norin 10 and Kokeshikomugi carried both Rht1 and Rht2. No tested variety carried Rht3. Some Norin varieties carrying Rht2 showed tall culms comparable to that of the rht tester line Chinese Spring. These results suggest that these varieties had a nullifier or modifier gene(s) or height promoting genes in the background controlling the height-reducing effect of Rht2. Conversely, six GA-responsive Norin varieties were as short as Akakomugi which carries the GA-responsive Rht genes, Rht8 and Rht9. The also seemed to carry a GA-responsive Rht gene or genes, and moreover all but one may carry gene(s) other than the Akakomugi genes. The origin of Rht1 and Rht2 of Norin 10 was speculated on the GA-response and Rht genotypes of its related varieties and landraces.     

Analysis of genic male sterility in Brassica oleracea

Summary The male sterility system MS-1 of Brassica oleracea was studied in order to elucidate if nucleo-cytoplasmic interactions determine this system. Crosses of male sterile MS-1 genotypes with heterozygous MS-5 genotypes gave fully fertile F₁ progenies. Selfing of seven F₁ plants resulted in five F₂ populations showing a 9:7 segregation ratio and two a 3:1 ratio for fertile and male sterile plants. Two F₂ progenies deviated from the expected 9:7 or 3:1 segregation ratios for fertile and male sterile plants. Thermosensitivity and distortion of the meiosis are suggested as the causal factors underlying the deviation of the segregation ratios. It was concluded that nuclear factors determine the male sterility in the MS-1 system, because the presence of a nucleocytoplasmic interaction in this system should have given only a 3:1 segregation ratio for fertile and male sterile plants in the F₂ generation.     

Effects of exogenous abscisic acid and gibberellic acid on pre-maturity α-amylase formation in wheat grains

Effects of in situ and in vitro applied abscisic acid (ABA) and gibberellic acid (GA₃) on pre-maturity ??-amylase (PMA) were investigated in glasshouse experiments under cool-temperature shock-induced and non-induced conditions using UK winter wheat varieties, Spark (low PMA susceptible) and Rialto (high PMA susceptible). In the in situ study, hormone solutions (ABA [100???M], GA₃ [50???M] and ABA?+?GA₃ [100?+?50???M]) were applied during mid-grain development to intact, developing grains of induced and non-induced plants. Alpha-amylase activity was measured in embryoless half-grains at maturity by a modified Phadebas assay. In the in situ study under non-induced conditions, applied ABA had no significant effect on ??-amylase in either variety whereas applied GA₃ significantly increased ??-amylase in Rialto, but not in Spark. In the in situ study under induced conditions, applied ABA and GA₃ produced no significant effect on ??-amylase in Spark; however in Rialto, applied ABA produced a small but significant decrease in ??-amylase whereas GA₃ significantly increased ??-amylase under induced conditions. In the in vitro study, spikes were harvested from induced/non-induced plants at 44/40?days after anthesis. Embryoless half-grains were incubated in hormone solutions at 25?°C and ??-amylase activity was measured. Results similar to the in situ study were observed in the in vitro study for both varieties and conditions, except GA₃ treatment which significantly increased ??-amylase under non-induced conditions in Spark. From these exogenous hormone studies, it appears that GA-response is a major factor during PMA induction by a cool-temperature shock whereas ABA-response seems to be of less importance.     

莲雾催花过程中叶片内源激素的变化

为探明莲雾产期调节的内在生理生化机理,以‘粉红种’莲雾为材料,研究莲雾反季节遮阳催花过程中叶片内源激素IAA、GA₃、ABA、ZT含量的动态变化和激素平衡（比值）的变化。结果表明：在遮阳处理后,叶片的IAA、ZT、GA₃、ABA含量均降低;喷施催花药剂后,IAA、ZT、ABA含量上升,GA₃含量下降。遮阳抑制了莲雾的营养生长。在花芽分化期,高水平的ABA、ZT,低水平的GA₃,IAA/ABA、GA₃/ABA、(IAA GA₃)/ABA低,ZT/GA₃高有利于莲雾成花。其中,高水平的ABA、GA₃/ABA低、ZT/GA₃高可能在莲雾的花芽分化中起主导作用。     

Factors affecting haploid production in wheat using the Hordeum bulbosum system. 3. Post-fertilization effects on embryo survival

Summary An investigation was made of factors affecting the frequency of development of fertilized ovules following wheat x Hordeum bulbosum hybridizations. Seed survival, a term used here to represent the relationship between seed set and fertilization, was shown to vary between the wheat genotypes tested, but was unrelated to the crossability genes present within the wheat genotype. A high seed survival was obtained in crosses involving the variety TH3929 and the single chromosome substitution line, Chinese Spring (Hope 5B). Chinese Spring showed a poor seed survival. Some influence of the H. bulbosum genotype on seed survival was detected. The two environmental factors studied, namely the post-pollination application of gibberellic acid and the ambient temperature, were both shown to affect seed survival. The application of gibberellic acid immediately after pollination or 1 and 2 days after pollination improved seed survival, although the extent of the response was dependent upon the number and timing of the application(s). A genotype-dependent response to the ambient temperature was observed, TH3929 showing a slight but insignificant reduction and Chinese Spring a significant improvement in seed survival with an increase in temperature from 20°C to 26°C. The physiological processes which may be influenced by GA and the ambient temperature and may thereby affect seed survival are discussed.     

A genetic analysis of the number of cells,length of cell,and gibberellic acid sensitivity in sugar beet and their relation to bolting mechanism

Summary Three series of full-sib progenies and parental inbred lines or populations were used to study the inheritance of the number of cells and length of cell along the hypocotyl length, and the response to gibberellic acid in young plants of sugar beet. Variance components were determined by using a factorial cross design. There were no significant estimates of non-additive genetic variation for either cellular characters. Both additive and non-additive genetic variance were responsible for the variability of stem length after GA3 treatment. It is concluded that additive type genes predominantly control the number of cells in all series. The length of cell had significant additive variances in most cases, but it was indeed markedly influenced by environmental factors. Number of cells (cell division rate) and GA3 sensitivity in young plants even before vernalization were related to bolting tendency. Bolting-susceptible genotypes generally expressed higher stem length in the response to added GA3 as compared to the bolting-resistant genotypes. However, some genotypes bolted easily in the field but reacted weakly to the GA3 treatment and vise versa. Genotypes that were susceptible to bolting and/or sensitive to GA3 had a specific range (intermediate) of cell number. There were, however, some genotypes containing an intermediate number of cells which demonstrated low bolting. These responded faintly to gibberellin treatment. The results suggest that several physiological requirements have to be fulfilled before bolting can occur, and that other plant characters interfere with the bolting phenomenon.     

Gibberellic acid as a gametocide for cole crops

Summary In preliminary and more elaborate trials GA_4/7 appeared to be an effective gametocide for Brussels sprouts, cabbage, cauliflower and kale.After repeated spraying of 10, 50 of 100 mg GA_4/7 per I isopropyl alcohol, during at least two weeks from the beginning of bolting, large numbers of male sterile flowers occurred during the early part of the flowering period. Many plants turned complete male sterile for periods up to 24 days.It is concluded that, depending on the type of cole crop, higher quantities of GA_4/7 or longer spraying periods are needed for permanent male sterility.     

A treatment to improve the vase life of cut tulips: Effects on tepal senescence, tepal abscission, leaf yellowing and stem elongation

The vase life of cut tulips (Tulipa spp.) is limited by a combination of leaf yellowing, tepal senescence, and tepal abscission. In many cultivars, moreover, high rates of stem elongation result in stem bending during vase life. In tests with the cvs. Apeldoorn and Frappant, stem bending could be prevented by treatment with ethylene or ethephon. However, these treatments resulted in poor flower opening. The ethephon treatment also resulted in precocious tepal abscission. The negative effect of ethephon on flower opening was overcome by a treatment with gibberellic acid (GA₃). GA₃ also somewhat delayed early leaf yellowing, but did not prevent the early tepal abscission induced by ethephon. The latter problem was overcome by a treatment with benzyladenine (BA). In addition, BA effectively delayed leaf yellowing and also delayed tepal senescence. However, BA produced browning of the lower stem end. This was prevented by the inclusion of calcium ions in the solution. The combination of chemicals (ethephon, GA₃, BA, and calcium ions) was similarly successful in a large number of other tulip cultivars tested. After adaptation of the concentrations the four chemicals were also effective if given as a pulse treatment shortly after harvest.