New insights into genotypic thermodependency of cytoplasmic male sterility for hybrid barley breeding

The cytoplasmic male sterility (CMS) system msm1 in barley is known to be thermosensitive, sometimes resulting in spontaneous fertility restoration in the absence of the corresponding restorer gene Rfm1. Here, we investigated genotypic differences concerning temperature sensitivity and the plant developmental stage at which elevated temperature induces spontaneous fertility restoration in three CMS mother lines. While one line stayed completely male sterile, a significantly higher fertility was observed in two lines after treatment from growth stage DC 41 until maturation. Microscopic analysis revealed that sterile anthers contained neither intact pollen, nor remains of aborted pollen grains, whereas pollen was visible in anthers of potentially fertile plants. We conclude that the barley CMS system affects anther and pollen development prior to meiosis. Elevated temperature during heading and flowering can lead to a spontaneous fertility restoration by reactivating pollen growth. Nevertheless, genotypic variation exists enabling the selection for stable CMS mother lines and the development of F₁ hybrids with high hybridity. As spontaneous fertility restoration due to environmental effects is difficult to phenotype, further investigations will focus on the development of molecular markers for marker‐assisted selection.     

Rapid phenotyping of adult plant resistance in barley (Hordeum vulgare) to leaf rust under controlled conditions

Breeding for adult plant resistance (APR) is currently impeded by the low frequency of annual field‐based testing and variable environmental conditions. We developed and implemented a greenhouse‐based methodology for the rapid phenotyping of APR to leaf rust in barley to improve the efficacy of gene discovery and cloning. We assessed the effects of temperature (18 and 23°C) and growth stage (1–5 weeks) on the expression of APR in the greenhouse using 28 barley genotypes with both known and uncharacterized APR. All lines were susceptible in week 1, while lines carrying Rph20 and several with uncharacterized resistance expressed resistance as early as week 2. In contrast, lines lacking Rph20 and carrying either Rph23 and/or Rph24 expressed resistance from week 4. Resistant phenotypes were clearest at 18°C. A subset of 16 of the 28 lines were assessed for leaf rust across multiple national and international field sites. The greenhouse screening data reported in this study were highly correlated to most of the field sites, indicating that they provide comparable data on APR phenotypes for screening purposes.     

Impact of Humidity on Temperature-Induced Grain Sterility in Rice (Oryza sativa L)

High temperature‐induced grain sterility in rice is becoming a serious problem in tropical rice‐growing ecosystems. We studied the mechanism of high temperature‐induced grain sterility of different rice (Oryza sativa L) cultivars at two relative humidity (RH) levels. Four varieties of Indica and Japonica rice were exposed to over 85 % RH and 60 % RH at 36/30 °C, 34/30 °C, 32/24 °C and 30/24 °C day/night air temperatures from late booting to maturity inside sunlit phytotrons. Increasing both air temperature and RH significantly increased spikelet sterility while high temperature‐induced sterility decreased significantly with decreasing RH. Neither Indica nor Japonica rice types were superior to the other in the response of their spikelets to increased air temperature and RH. Increased spikelet sterility was due to increased pollen grain sterility which reduced deposition of viable pollen grains on stigma. Reduction in sterility with decreased RH was more due to decreased spikelet temperature than to air temperature. Thus the impact of RH should be considered when interpreting the effect of high temperature on grain sterility. Spikelet fertility was curvilinearly related to spikelet temperature. Grain sterility increased when spikelet temperature increased over 30 °C while it became completely sterile at 36 °C. The ability of a variety to decrease its spikelet temperature with decreasing RH could be considered as avoidance while the variability in spikelet sterility among varieties at a given spikelet temperature could be considered as true tolerance.     

Sorghum can Compensate for Chilling-Induced Grain Loss

Chilling during male gametophyte development in sorghum (Sorghum bicolor L. Moench) inhibits development of microspores, causing male sterility. The aim of this study was to assess the effects of night chilling on yield components in sorghum. This study identified and employed collar distance as a morphological marker of anther development following chilling. Two cultivars Buster and Bonus, were subjected to three temperature regimes (25/20 °C, 25/12 °C and 25/8 °C) for five consecutive nights at 0 cm collar distance, which corresponds to the meiotic stage of anther development. Pollen viability, grain number and harvest index were reduced in both cultivars at 12 and 8 °C night temperatures. Total grain weight of cultivar Bonus was not reduced as much as Buster at 12 °C because of the ability of Bonus to increase individual grain weight when grain number was low. This work further elucidates the mechanism and genetic potential of chilling‐induced yield compensation for developing sorghum cultivars that are better adapted to low night temperatures.     

Maternal inheritance of male sterility in the progeny of a natural hybrid between Cajanus lineatus and C. cajan

Adoption of pigeonpea hybrids in central and southern India is showing high impact with on‐farm yield advantages of >30%. The hybrid pigeonpea technology, the first in any legume crop, is based on a cytoplasmic‐nuclear male‐sterility (CMS) system. For a long‐term sustainability of hybrid programme, it is imperative that both nuclear diversity and cytoplasmic diversity are maintained among hybrid parents. In this context, a continuous search for new CMS‐inducing cytoplasms is necessary. This paper reports detection of maternal inheritance of male sterility in the progeny derived from a natural hybrid between a wild relative [Cajanus lineatus (W. & A.) Maesen comb. nov.] of pigeonpea and an unknown pigeonpea [Cajanus cajan (L.) Millsp.] genotype. In the present study, the male sterility was maintained up to BC₇F₁ generation by an advanced breeding pigeonpea line ICPL 99044. This male sterility inducing cytoplasm of C. lineatus was tagged as A₆. In future, this CMS genetic stock can be used to develop a range of new pigeonpea hybrids with high yield and adaptation.     

Variations in chlorosis and potential usefulness of alloplasmic Brassica rapa with the cytoplasm of male sterile Brassica juncea

To select superior seed parents for vegetable hybrid seed production, we conducted interspecific crosses between male sterile Brassica juncea (2n = 36, AABB) and eight inbred lines of Brassica rapa (2n = 20, AA). Alloplasmic lines of B. rapa with the cytoplasm of B. juncea were developed from B. juncea × B. rapa hybrids by repeated backcrossing using B. rapa as the recurrent male parent until the BC₃ generation. Seed fertility, male sterility and chlorophyll content were investigated in these plants cultivated under four different temperature conditions (5, 10, 12 and 20°C). At 10°C, the alloplasmic lines of B. rapa with the cytoplasm of B. juncea were male sterile with partly chlorotic leaves. The alloplasmic B. rapa had lower chlorophyll a, chlorophyll b and carotenoid contents than those of the original B. rapa. The leaves recovered from chlorosis when the plants were cultivated at 20°C. An alloplasmic line of B. rapa (A6) is available as a seed parent for vegetable hybrid seed production and contributes seed fertility, slight chlorosis and stable male sterility.     

Toward pollen sterility in German chamomile (Matricaria chamomilla)–Conventional breeding approaches of cytoplasmic/genic male sterility and chemical emasculation

Pollen sterility (PS) in mother lines is crucial for hybrid crossings in plant breeding. 21 di- (2x) and tetraploid (4x) accessions of the hermaphroditic chamomile were screened for spontaneous, thermic or gametocide induced PS. Two crossing trials with the diploid accessions 'Bona' (BON) and 'Hungary 2' (HUN2) should specify maternally inherited cytoplasmic male sterility (CMS). Progeny thereof showed increased PS. BON as mother (BH) entailed means of 30 and 22% PS, versus 8% and 20% after reciprocal crossings. Backcrosses of BH with HUN2 in paternal and maternal direction revealed increasing mean PS (24% and 32%), without verification of CMS. A split plot design of 100 individuals treated with four chemical gametocides analyzed PS, seed set, germination rate and whole plant damage. Tri-iodobenzoic acid caused the highest PS (43%). Progeny of two plants (2x, 4x) with degenerated anthers without pollen, showed 100% PS either in tetraploid F₁ and F₂ (4% and 11%) or in diploid F₂ and F₃ (26% and 23%). High temperature (30/28°C day/night) decreased the male fertility to some extent.     

Use of barley seed vigour to discriminate drought and cold tolerance in crop years with high seed vigour and low trait variation

Seed vigour is a precondition for early and homogenous field emergence of barley, in addition to effective malting. This study aimed to assess the selection of barley varieties by using seed vigour as the indicator. Seed vigour of barley (quantified as the germination percentage) was evaluated under drought (?0.2 MPa) and temperature stress (10°C). At two locations over a 3‐year period, 1 population of 133 Derkado × B83‐12/21/5 doubled haploid (DH) lines (and parents) was evaluated for seed vigour, of which 108 DH lines were assessed for three malting parameters. The relatively high values of vigour during the 3‐year period (overall average 94–95%) probably impeded high variations in genetic potential. A total of 27 DH lines of the 133 evaluated showed transgression for vigour (up to 98%) in comparison with the parents (Derkado: 96%; B83: 92%). In conclusion, caution should be applied when selecting for seed vigour, even in good crop years with high levels of seed vigour and low trait variations. Such selection might improve vigour, particularly in crop years with unsuitable weather conditions.     

Inheritance and allelic relationships of fertility restoration genes for seven new sources of male-sterile cytoplasm in sunflower

The inheritance of fertility restoration of six mitomycin C and streptomycin‐induced cytoplasmic male‐sterile (cms) mutants and one cms line derived from Native American cultivar PI 432513 in sunflower was evaluated. These seven new cms sources were also compared with the commercially used cms PET1 (Helianthus petiolaris Nutt.) cytoplasm, using USDA inbred lines with restoration genes (Rf₁) specific for cms PET1 and new restoration lines identified for cms PI 432513. Restoration genes for cms PI 432513 were found in ‘Armavir’, VNIIMK, P21 and male‐fertile (MF) plants of PI 432513. F₂ and F₃ segregation ratios of crosses between cms PI 432513 and these restoration sources indicated a single dominant gene controlled fertility restoration. Progenies of cms PI 432513 testcrossed with F₁’s of half‐diallel crosses among the respective four homozygous restoration lines and RHA 274 suggested that the restoration genes of RHA 274, VNIIMK, P21 and PI 432513 were at the same locus. Restoration genes from VNIIMK, P21 and PI 432513 satisfactorily restored pollen stainability in the heterozygous condition. A very weak expression of the Rf gene in ‘Armavir’ was observed in the heterozygous condition. Fertility restoration capability of these genes for the six mutant cms HA 89 and cms HA 89 (in PET1 cytoplasm) was observed. The mutant cms HA 89 lines were also restored completely by RHA 266, RHA 274, RHA 280 and RHA 296, and F₂’s segregation ratios indicated single dominant gene control, implying a common cytoplasmic male sterility in all lines. F₁’s of half‐diallel crosses among RHA 266, RHA 273, RHA 274, RHA 280 and RHA 296 were testcrossed onto the cms lines, and their all MF progenies among lines, except RHA 280, confirmed that fertility restoration was controlled by a single Rf₁ gene locus. The restoration gene in confection line RHA 280, namely Rf₃, was at a different locus than Rf₁ and was equally capable of restoring all the cms lines. Cms HA 89 mutants and cms PI 432513 are in H. annuus cytoplasm, and are agronomically equal in hybrid performance to the cms PET1 used in commercial sunflower hybrids. These new cms lines will provide immediate alternative cms sources for reducing the genetic vulnerability resulting from the exclusive use of the single cms source PET1 in sunflower hybrid production.     

Expression of thermosensitive genic male sterility in rice under varying temperature situations

Response of thermosensitive genic male sterility in rice to varying temperature situations was studied by using four TGMS lines. In three sets of maximum, minimum and their combined temperatures, it was observed that maximum temperature played a predominant role in influencing sterility/fertility of TGMS lines under the combined regimes. Expression of a TGMS gene was found to be influenced by the genetic background of the recipient lines. Exposure for more than 8 hours of 32 ^°C was found necessary to induce complete male sterility in indica TGMS lines: IR68945-4-33-4-14 and IR68949-11-5-31 while, more than 4 hours of such exposure was enough to induce sterility in case of their japonica donor Norin PL 12. Sudden interruption with 27 ^°C even for 2 hours under the sterility inducing regime of 32/24 ^°C could induce partial fertility in the line IR68945-4-33-4-14. However, the line ID 24 remained completely sterile even with 10 hours of interruption with 27 ^°C. An interruption with lower temperature of 27 ^°C for 4 hours for even one day induced partial fertility in IR68945-4-33-4-14. The period of four to eight days after panicle initiation was the stage most sensitive to temperature variations. Hybrid rice breeders need to develop numerous genetically diverse TGMS lines, which possess critical sterility inducing temperature of 28 ^°C and are not affected by sudden interruptions with a lower temperature for few hours daily for a couple of days. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetics, fertility behaviour and molecular marker analysis of a new TGMS line, TS6, in rice

The thermosensitive genic male sterility (TGMS) system has great potential for revolutionizing hybrid rice production through simple, less expensive and more efficient seed production technology. For the successful utilization of this novel male sterility system, knowledge of the breeding and fertility behaviour of a TGMS line is essential. In this study, the fertility transformation behaviour, the critical fertility and sterility temperatures and the mode of inheritance of male sterility were studied for a new TGMS line, TS6, identified at Tamil Nadu Agricultural University, Coimbatore, India. The pollen and spikelet fertilities recorded on plants raised at fortnightly intervals revealed that this line was completely sterile for 78 consecutive days (35/22 to 32/23°C, maximum/minimum temperatures) and reverted to fertile when the temperature was 30/18°C. It remained fertile continuously for 69 days and the maximum pollen and spikelet fertilities recorded were 75 and 70%, respectively. The fertility was highly influenced by daily maximum temperature followed by average and minimum temperatures. It was not influenced by relative humidity, sunshine hours or photoperiod. The critical temperature inducing sterility and fertility was 26.7 and 25.5°C, respectively. The male sterility in TS6 was inherited as a monogenic recessive in the F₂ and BC₁ populations of TS6 × MRST9 as well as TS6 × IR68281B. Using bulked segregant analysis on an F₂ population of TS6 × MRST9, an RAPD marker, OPC05₂₉₆₂, was identified to be associated with TGMS in TS6.     

Growth and Physiological Responses of Quinoa to Drought and Temperature Stress

Quinoa (Chenopodium quinoa Willd.), traditionally called the mother of grains, has the potential to grow under high temperatures and drought, tolerating levels regarded as stresses in other crop species. A pot experiment was conducted in a climate chamber to investigate the potential of quinoa tolerance to increasing drought and temperature. Quinoa plants were subjected to three irrigation and two temperature regimes. At low temperature, the day/night climate chamber temperature was maintained at 18/8 °C and 25/20 °C for high temperature throughout the treatment period. The irrigation treatments were full irrigation (FI), deficit irrigation (DI) and alternate root‐zone drying (ARD). FI plants were irrigated daily to the level of the pot's water‐holding capacity. In DI and ARD, 70 % water of FI was applied either to the whole pot or to one side of the pot alternating, respectively. The results indicated that plant height and shoot dry weight significantly decreased by ARD and DI compared to FI treatment both at low and at high temperatures. However, plants in ARD treatment showed significantly higher plant height and shoot dry weight compared to DI especially at higher temperature, which is linked to increased xylem ion content. Higher quinoa plant growth in ARD was associated with increase in water‐use efficiency (WUE_i) due to higher abscisic acid concentration and higher nutrient contents compared to DI. From results, it can be concluded that quinoa plant growth is favoured by high temperature (25/20 °C) and ARD is an effective irrigation strategy to increase WUE in drought prone areas.     

Genetic Control of Temperature-Sensitivity of Cytoplasmic Male Sterility (cms) in Chives (Allium schoenoprasum L.)

In earlier experiments with chives (Allium schoenoprasum L.) temperature-sensitive and temperature-insensitive cms plants could be selected. To obtain information about the site and nature of the genetic factor(s) responsible for the temperature sensitivity of the cms, temperature-insensitive and temperauire-sensitive cms plants were crossed with maintainers as well as with one another. In the progenies each genotype was cloned into two clone members, and these were examined concerning their male sterility under normal (20 °C/14 °C, day/night) as well as under constantly high temperatures (24 °C/ 24 °C). The results indicate that the temperature-sensitivity of the cms is controlled by a dominant nuclear gene “T”. This acts like a restorer gene at high temperatures and is ineffective at normal temperatures. Accordingly, important conclusions can be drawn with regard to the selection of temperature-insensitive cms-lines as well as to the propagation of the cmi-lines. The results are discussed in connection with the mitochondrial polypeptides probably responsible for the formation of the cms.     

Development of cytoplasmic-genic male sterility in safflower

An interspecific cross was made between Carthamaus oxyacantha and the cultivated species C. tinctorius to develop a cytoplasmic‐genic male sterility (CMS) system in safflower. C. oxyacantha was the donor of sterile cytoplasm. The 3: 1 segregation pattern observed in BC₁F₂ suggested single gene control with dominance of male‐fertility over male‐sterility. The information obtained from crossing male sterile X male fertile plants in BC₁F₃ and BC₁F₄ generations showed statistically significant single gene (1: 1) segregation for male sterility vs. male fertility. The results demonstrated that C. tinctorius possesses a nuclear fertility restorer gene and that a single dominant allele restored fertility (Rf) in progeny carrying CMS cytoplasm of C. oxyacantha. Male sterility occurred with the homozygous recessive condition (rfrf) in a sterile C. oxyacantha cytoplasm background and not in the normal cytoplasm of C. tinctorius. The genetic background of different restorer lines of C. tinctorius having normal cytoplasm did not effect fertility restoration. The absence of male sterile plants in C. tinctorius populations ruled out the possibility of genetic male sterility. Normal meiosis in F₁ and BC₁F₂ ruled out a cytogenetic basis for the occurrence of male sterility.     

Effect of high temperature on pollen morphology,plant growth and seed yield in quinoa (Chenopodium quinoa Willd.)

Quinoa (Chenopodium quinoa Willd.) has gained considerable attention worldwide during the past decade due to its nutritional and health benefits. However, its susceptibility to high temperatures has been reported as a serious obstacle to its global production. The objective of this study was to evaluate quinoa growth and pollen morphology in response to high temperatures. Pollen morphology and viability, plant growth and seed set, and several physiological parameters were measured at anthesis in two genotypes of quinoa subjected to day/night temperatures of 22/16°C as a control treatment and 40/24°C as the heat stress treatment. Our results showed that heat stress reduced the pollen viability between 30% and 70%. Although no visible morphological differences were observed on the surface of the pollen between the heat‐stressed and non‐heat‐stressed treatments, the pollen wall (intine and extine) thickness increased due to heat stress. High temperature did not affect seed yield, seed size and leaf greenness. On the other hand, high temperature improved the rate of photosynthesis. We found that quinoa has a high plasticity in response to high temperature, though pollen viability and pollen wall structure were affected by high temperatures in anthesis stage. This study is also the first report of quinoa pollen being trinucleate.     

Development of thermo-sensitive cytoplasmic male sterile (TCMS) lines of wheat characterized by complete male sterility at lower-temperatures and partially restored fertility at higher-temperatures

A ‘‘two-line hybrid system’’ was developed using thermo-sensitive cytoplasmic male sterility (TCMS) from Aegilops kotschyi and is proposed as a means for seed multiplication and production of hybrid wheat. The TCMS line is crossed to a pollinator line during the normal wheat-growing season for production of hybrid wheat seeds, whereas the TCMS line is maintained through self-pollination in an environment with high temperatures. The male sterile wheat line KTP3314A, containing Ae. kotschyi cytoplasm and the short arm of chromosome 1B from Triticum spelta, was studied in different temperature regimes. KTP3314A was completely male sterile under normal wheat growing temperatures (<18 °C). Sterility of KTP3314A was partially restored when plants were grown at temperatures above 20 °C during growth stages 45–52. Male fertility was not correlated with day-length. A series of TCMS lines derived from KTP3314A showed similar environmental responses and performed well in hybrid wheat production at lower temperatures. Fertility of these TCMS lines at higher temperatures makes it possible to produce large quantities of male sterile parental seeds. Based on these findings, application of the TCMS-dependent two-line system in hybrid wheat breeding appears to have a promising future for hybrid wheat production.     

Establishing Cytoplasmic Male Sterility in Brassica napus by Mitochondrial Recombination with B. tournefortii

Fusion experiments between B. napus and X-ray-treated B. tournefortii protoplasts were carried out to develop cytoplasmic male sterility (ems) in B. napus. From the regenerants, six lines containing male sterile plants were selected; five lines segregated for male sterility, but one line (25–143) was completely male-sterile from the beginning. Molecular analyses of mitochondrial (mt) and chloroplast (cp) DNA of B. napus, B. tournefortii, B. juncea and cms juncea indicated that the original cytoplasmic donor of the cms juncea-system in B. napus was a B. tournefortii form, while the B. napus genotype used for the fusion experiments had a B. campestris cytoplasm. By analysis (it regenerated plants, line 25–143 was identified as possessing mt-DNA recombined between B. campestris and B. tournefortii. with the major part derived from B. campestris. No differences were detected between epDNAs from H. campestris and from line 25—143. The other five lines were similar to B. campestris with all the probes used. The low frequency of sterile lines from the fusion experiments and the inheritance of the cms in segregating progenies are both discussed.     

甘蓝型油菜胞质雄性不育系22A的遗传研究

甘蓝型双低油菜CMS22A是河南省农科院棉油所利用384A中的雄性不育源、采用单株成对连续回交的方法育成的新型双低油菜细胞质雄性不育系。其突出表现为雄性不育彻底且遗传稳定,配合力高,抗病抗倒,品质优良。对CMS22A育性遗传规律研究表明,该不育系属典型的败药型雄性不育系,其雄性不育的遗传受不育细胞质(S)和一对隐性核不育主效基因(rr)的共同控制,属孢子体不育,基因型为S(rr)。     

Effects of Mild Temperature Stress on Grain Quality and Root and Straw Nitrogen Concentration in Malting Barley Cultivars

Abstract It is a challenge to obtain the appropriate protein concentration in cereals for the intended end‐use. This study examined ambient temperature effects on two spring malting barley cultivars (Henley and Tipple) grown in soil or in solution culture with controlled nitrogen supply in daylight chambers with low temperature (day 18 °C, night 12 °C), and high temperature (23 °C/17 °C) to/after anthesis. In soil‐grown plants, high temperature to anthesis resulted in higher grain nitrogen amount (GNA), grain nitrogen concentration (GNC) and straw nitrogen concentration (SNC). In plants grown in solution, high temperature to anthesis resulted in lower GNA and higher GNC. A temperature rise of 1 °C during the growing period in solution cultivation increased GNC, root nitrogen concentration (RNC) and SNC, by 1.20, 1.35 and 0.33 mg g^?1, respectively. In solution culture, GNC was positively correlated with RNC and SNC (P < 0.01). Cv. Henley had higher GNC but lower SNC than cv. Tipple. Cv. Henley was more stable in grain size and cv. Tipple in GNC. The results showed that temperature has a direct effect on GNC. Accounting for temperature fluctuations up to the latest possible nitrogen fertilisation occasion can therefore help when deciding appropriate nitrogen supply for intended end‐use.     

Ethyloxanilates as specific male gametocides for wheat (Triticum aestivum L.)

Induction of male sterility by deployment of male gametocides holds immense potential in heterosis breeding of wheat. The efficacy of a new class of male gametocide for wheat (Triticum aestivum L.) is described: ethyloxanilates, the most active example of this class being ethyl 4‐fluorooxanilate (E4FO). E4FO induces male sterility, specifically, without detectable effects on various agronomic features and female fertility. The plants sprayed once with 0.15% E4FO exhibited 100% pollen and floret sterility without causing a significant reduction in total yield. E4FO was screened on 29 genotypes of wheat at 0.15% test concentration and was observed to induce 99.76 ± 0.37% male sterility. Thirteen F₁‐hybrids of wheat were produced using the gametocide in Winter 2000–2001 and were evaluated for their agronomic performance in Winter 2001–2002. The cross combinations viz., lines WR 544 × HW 2046 and HW 2044 × WR 956 have outperformed their respective better parents by 48.17% and 23.42% in grain yield/plant and thus have potential as hybrids.