The influence of anther cold pretreatments and donor plant genotypes on in vitro androgenesis in wheat (Triticum aestivum L.)

Cold pretreatments applied to excised anthers in liquid potato 2 medium proved to be unnecessary. Generally, cold pretreatments inhibited anther response and productivity as the duration was lengthened or as the pretreatment temperature was lowered. There were significant differences in response attributed to the anther donor genotype. Green and albino plants as well as roots only have been regenerated from the spring wheat cultivars Sinton, Neepawa, Pitic 62 and DW 50. Most plants were haploid.     

Factors inducing regeneration response in oat (Avena sativa L.) anther culture

The efficiency of embryogenesis of anther culture was compared using four cultivars of oat (Avena sativa L.): ‘Akt’, ‘Bingo’, ‘Bajka’, and ‘Chwat’. Despite the high resistance of oat to the process of androgenesis, all tested cultivars produced embryo-like structures and only two of them, ‘Akt’ and ‘Chwat’, produced fertile doubled haploid plants. A strong cultivar dependency was observed during induction of androgenesis. Further, cold pretreatment together with high temperature shock enhanced the efficiency of this technique. The highest number of embryo-like structures and haploid plants was obtained from cv. ‘Chwat’ (3.6% and 0.8%, respectively). Embryo-like structure formation also depended on the distance from the base of the flag leaf to the penultimate leaf of the panicle. Most of them were observed on anthers harvested from panicles of which the distance from the base of the flag leaf to the penultimate leaf was less than 4 cm. The presence of the induction medium supplemented with different plant growth regulators was essential for the induction of embryo-like structures but did not increase the production of haploid plants and doubled haploid lines. The highest number of embryo-like structures and plants was obtained on W14 medium with the addition of 2.0 mg/dm³ 2,4-dichlorophenoxyacetic acid and 0.5 mg/dm³ kinetin (2.7%). The low haploid plant regeneration rate (from 0.03 to 0.05%) still limits the practical application of anther culture for the production of doubled haploid lines in oat.

     

低温和PEG预处理对小麦愈伤组织形成及IAA氧化的影响

研究了低温和PEG预处理对小麦成熟胚愈伤组织形成及IAA氧化的影响。结果表明,低温和PEG预处理促进两种小麦成熟胚愈伤组织形成,但对其IAA氧化的影响不同;其中低温引起‘郑引1号’IAA氧化酶活性降低,但对‘保7059’的影响不大;而PEG使‘保7059’的IAA氧化酶活性增加,但对‘郑引1号’无明显影响;两种处理均使两种小麦IAA过氧化物酶活性降低。     

Induction of in vitro androgenesis in anther and isolated microspore culture of different spelt wheat (<Emphasis Type="Italic">Triticum spelta</Emphasis> L.) genotypes

This is the first report on isolated microspore culture—derived spelt wheat. The efficiency of anther- and isolated microspore was compared using four genotypes (‘Franckenkorn’, ‘GK Fehér’, ‘Mv Martongold’, ‘Oberkulmer Rotkorn’). In anther culture, genotype dependency was observed, and cold pre-treatment enhanced the efficiency of the method. In isolated microspore culture, the ovary co-culture supported the development of embryo-like structures. The presence of growth regulators (0.5 mg/l 2,4-D and 0.5 mg/l kinetin) were not essential for the induction of androgenesis, but these increased the production of embryo-like structures, green and albino plantlets. The low plant regeneration rate and high number of albinos hinder the practical application of isolated microspore culture while anther culture was efficient for in vitro green plantlets production in spelt wheat. The mean of green plantlets production was 41.45/100 anthers (from 20.93 to 83.07 depending on genotype). The phenomenon of albinism was mitigated in anther culture (3.48 albinos/100 anthers). Altogether, 1720 anther culture—derived green plantlets were produced from the four genotypes.     

Cold pretreatment to enhance green plant regeneration from rye anther culture

Cold pretreatment of detached tillers of rye, Secale cereale, was tested under two light regimes. The ryes included two spring and two winter cultivars. Significant increases in green plant regeneration were recorded in each experiment when cold pretreatments of two to four weeks were applied. Dim light during the stress period improved green plant regeneration for two of the four cultivars tested. The highest regeneration rate, 30.6 green plants per 100 plated anthers, was reached following three weeks at +4 °C under dim light, for spring rye Jo02. Starvation stress applied to plated anthers in mannitol medium suppressed anther response. This revised version was published online in June 2006 with corrections to the Cover Date.     

Doubled haploid production via anther culture in annual,winter type of caraway (<Emphasis Type="Italic">Carum carvi</Emphasis> L.)

Caraway (Carum carvi L.) is a traditional medicinal and spice cross-pollinated plant species. Although in vitro techniques are recently extensively applied in plant breeding programmes, these are not commonly utilized in caraway. Therefore, based on the protocol for anther culture in carrot (Daucus carota L., a closely related species of caraway in Daucaceae family), in vitro androgenesis in caraway has been studied with the aim to produce completely homozygous inbred lines. Various induction conditions, such as temperature pretreatments, carbon sources and combination of growth regulators in a culture medium as well as the effect of genotype on in vitro androgenesis were examined. Ten breeding lines of winter caraway representing third generation of forced (artificial) self-pollination were used as donor plant material. Cultured anthers produced embryogenic calli, and subsequently two types of regenerated plants were obtained, namely haploids with evident microspore origin, and diploids which may represent somatic (anther wall) regenerants or spontaneous doubled haploids. The ploidy status of regenerated plants was determined by flow cytometry. This is the first report on androgenic doubled haploid production in caraway.     

Production of total and stainable pollen grains in Olea europaea L

The production of the total and stainable number of pollen grains per flower and per inflorescence of 13 olive cultivars (Olea europaea L.) has been investigated. Grains stainability was evaluated by using the acetic carmine staining method whereas the production of pollen grains was calculated by using a Bürker haemocytometer. All pollen characteristics taken into account varied significantly among the cultivars. The number of flowers per inflorescence ranged from a maximum of 31.3 (‘Cipressino’) to a minimum of 13.0 (‘Leccino’), the number of pollen grains per anther ranged from 123?346 (‘Arbequina’) to 40?975 (‘Sant'Agostino’), and the pollen stainability from 97.6% (‘Cipressino’) to 48.0% (‘Cellina di Nardò’). The number of stainable pollen grains per inflorescence averaged 2?559?512, ranging from a maximum of 3?913?678 (‘Nociara’) to a minimum of 940?143 (‘Sant'Agostino’). All parameters were positively correlated, whereas a linear negative correlation was found between stainability and total pollen grains both per anther and per inflorescence.     

Factors affecting somatic embryogenesis in anther cultures of Chinese pink <Emphasis Type="Italic">(Dianthus chinensis</Emphasis> L<Emphasis Type="Italic">.)</Emphasis>

In this study, we aimed to maximize the rates of somatic embryogenesis achievable in anther cultures of Chinese pink (Dianthus chinensis L.) (2n = 2x = 30). The genotype of the donor plant was found to be a major factor in determining the success rate. Conditions imposed during anther culture (notably medium composition and light conditions) and pretreatments (namely, cold, heat, and mannitol incubations) were also found to influence somatic embryo induction. For example, the highest levels of embryogenic callus induction were achieved when the donor buds had been cold pretreated and the subsequent anther culture was maintained in darkness. Furthermore, there appeared to be an interaction of genotype with culture conditions. Thus, in cultures of the cultivar (cv.) ‘Carpet’, the highest rates of embryogenesis were obtained when the anthers had received a 5-d heat-shock, but such a thermal treatment did not generally produce a significant effect. Likewise, a 3-d mannitol pretreatment was optimal only for the cross-hybrid line ‘HC’. Assessment of the ploidy of the plants regenerated from the anther cultures revealed both diploid and tetraploid plants. Histological and cytological observations showed that all of these (both from n-pollen and 2n-pollen lines) derived from anther wall cells. Spontaneous chromosome doubling was inferred to have occurred during the embryogenic callus culture period.     

Production and conversion of haploid embryos in chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) anther cultures using high 2,4-D and silver nitrate containing media

Due to recalcitrant nature of chickpea (Cicer arietinum L.) to androgenesis, the production of double haploid plants has been only reported by Grewal et al. (Plant Cell Rep 28:1289–1299, 2009) using some physical stresses such as anther centrifugation and electrical shock. In the present study, we successfully obtained haploid plants from cultured anthers of two chickpea cultivars, Bivanij and Arman, using high 2,4-D and silver nitrate containing media without applying of these time and labor consuming stresses. For induction of androgenesis, different concentrations of 2, 4-D (0, 2, 5 and 10 mg/l) and silver nitrate (0, 5, 10, 15, 25 and 50 mg/l) were used in embryo development medium. In Bivanij cultivar, anther induction medium containing 10 mg/l 2,4-D and 15 mg/l silver nitrate produced the highest number of embryos (0.188) and regenerated plants (0.1) per each cultured anther, while the highest frequencies of embryos (0.1) and regenerated plants (0.075 and 0.063) were obtained from Arman cultivar when 10 mg/l 2,4-D was combined with 15 and 50 mg/l silver nitrate in anther culture medium, respectively. In second part of this study, different cold (4 °C for 4 and 7 days) and heat (30 °C for 10 days, 32 °C for 2 days and 35 °C for 8 h) pretreatments were applied on cultured anthers of Bivanij cultivar. Incubation of cultured anthers at 32 °C for 2 days significantly enhanced the rate of embryo formation up to 0.222 embryos per each anther, while the highest number of regenerated plants/anther (0.0332) was obtained when cold treated anthers at 4 °C for 7 days incubated at 30 °C for 10 days. Taken together, these results provide a good basis for large-scale generation of DH plants in this important legume species.     

Reciprocal effects in anther cultures of wheat hybrids

This study was conducted to determine the reciprocal effects for anther culture response in wheat (Triticum aestivum L.) using a set of 4 × 4 full diallel crosses. Both reciprocal and nuclear genetic effects were highly significant for anther culture response and useful for selection and breeding purposes. General combining ability (GCA) effects were predominant for all investigated anther culture traits. Also, significant differences for specific combining ability (SCA) effects were detected between reciprocal crosses. Although significant reciprocal differences for responding anther, callus number and green plant regeneration were recorded in some reciprocal crosses, there were no significant reciprocal differences for albino plant regeneration. The use of one parent as male or female could lead to change at the production of green plants from the F₁ hybrids and screening of inbred lines for response to anther culture, without reciprocal effects, could decrease the utilization of breeding material.     

Morphological markers to correlate bud and anther development with microsporogenesis and microgametogenesis in pepper (Capsicum annuum L.)

The identification of microspores or pollen grains at particular developmental stages during microsporogenesis or microgamentogenesis is an important step for different basic and applied purposes. Among them, the most relevant example from a biotechnological perspective is the production of androgenic doubled haploids. For this and other techniques, precise, fast, easy and reliable criteria to identify flower buds carrying microspores or pollen at particular stages are essential. In anthocyanin-producing pepper types, the particularities of flower development allow for the identification of several morphological markers potentially useful as criteria for such an identification. In this work, our aim was to determine the easiest and more accurate criterion to correlate visible, measurable traits of bud and anther development with each of the individual stages of microsporogenesis and microgametogenesis. For this, we used three Spanish sweet pepper F1 hybrids (‘Herminio’, ‘Gacela’ and ‘Águila’). We analyzed and discussed the accuracy and practical usefulness of using anther length, bud length, anther purple pigmentation and the ratio between calyx length and bud length (calyx/bud ratio) as predictors of individual microspore/pollen developmental stages. According to our results, we propose a combination of calyx/bud ratio and anther pigmentation as an easy, fast and accurate criterion potentially applicable to anthocyanin-producing pepper cultivars to determine their particular markers.     

A comparative study of genotypic and environmental response to androgenesis in Nicotiana rustica

Summary A total of six genotypes of Nicotiana rustica comprising the two F₁'s (V₂ × V₁₂ and V₁ × V₅) and their parents were evaluated for their efficiency in haploid production. Excised immature flower buds with pollen at late uninucleate to early binucleate stage were pretreated for 21 days at 5 ° or 7 °C, or for 15 days at 9 °C before culturing on Nitsch's medium+ 0.1 mg/l NAA. The effects of genotype, pretreatment and their interaction were tested on anther response, anther productivity and days to first plantlet formation. Highly significant genotype X pretreatment interaction and differences between genotypes were observed for all three characters. Significant differences between pretreatments were observed for anther productivity only. The performance of V₁₂ both in respect of anther productivity and response was highest whereas that of V₅ was the lowest. Analysis of variance showed that a simple additive genetic model was not adequate to explain the above variation due to significant additive genetic and dominance interactions with the pretreatment.     

Response to cadmium in higher plants

The paper summarizes present knowledge in the field of higher plant responses to cadmium, an important environmental pollutant. The principal mechanisms reviewed here include phytochelatin-based sequestration and compartmentalization processes, as well as additional defense mechanisms, based on cell wall immobilization, plasma membrane exclusion, stress proteins, stress ethylene, peroxidases, metallothioneins, etc. An analysis of data taken from the international literature has been carried out, in order to highlight possible ‘qualitative’ and ‘quantitative’ differences in the response of wild-type (non-tolerant) plants to chronic and acute cadmium stress. The dose-response relationships indicate that plant response to low and high cadmium level exposures is a very complex phenomenon indeed: cadmium evokes a number of parallel and/or consecutive events at molecular, physiological and morphological levels. We propose that, above all in response to acute cadmium stress, various mechanisms might operate both in an additive and in a potentiating way. Thus, a holistic and integrated approach seems to be necessary in the study of the response of higher plants to cadmium. This multi-component model, which we would call ‘fan-shaped’ response, may accord with the Selyean ‘general adaptation syndrome’ hypothesis. While cadmium detoxification is a complex phenomenon, probably under polygenic control, cadmium ‘real’ tolerance—found in mine plants or in plant systems artificially grown under long-term selection pressure, exposed to high levels of cadmium—seems to be a simpler phenomenon, possibly involving only monogenic/oligogenic control. We conclude that, following a ‘pyramidal’ model, (adaptive) tolerance is supported by (constitutive) detoxification mechanisms, which in turn rely on (constitutive) homeostatic processes. The shift between homeostasis and ‘fan-shaped’ response can be rapid and involve quick changes in (poly)gene expression. Differently, the slow shift from ‘fan-shaped’ response to ‘real’ cadmium tolerance is caused and affected by long-term selection pressure, which may increase the frequency (and promote the expression) of one or a few tolerance gene(s).     

The response of anther culture to culture temperature varies with growth conditions of anther-donor plants

The response of anther culture to culture temperature was tested using anther from wheat (Triticum aestivumL.)(3 spring varieties and 1 semi-winter variety) grown under two different conditions, in the field in spring and summer and in the greenhouse in winter and spring in Beijing. The results showed that the most suitable culture temperature for anther culture of field-grown materials was about 2°C higher than that of the greenhouse-grown materials, no matter what genotype was used. For example, in anther culture of the variety Jinghong 5, the highest yield of green pollen plantlets appeared at a culture temperature of 30°C when the anther-donor plants had been grown in the greenhouse, but was at 32°C when the donor plants were grown in the field. It was noticed that the anther of the greenhouse-grown material did not develop as well as those of the field-grown material.     

Differential colonization of wheat cultivars by two biotypes of Russian wheat aphid （Homoptera： Aphididae）

Susceptible and resistance wheat cultivars, Triticum aestivum L, were presented to two biotypes of Russian wheat aphid, Diuraphis noxia （Mordvilko）, in multiple choice tests to assay their relative acceptability as host plants. Both apterae （third and fourth instars） and alate adults were offered plants at the two-leaf stage in different cultivar combinations at 22±1℃ and 16：8 （L： D） hour photoperiod. Apterae were released from Petri dishes in the center of a circle of test plants, whereas alatae dispersed from a mature aphid colony to settle on plants arranged in rows. Both alatae and apterous nymphs of both biotypes readily colonized all cultivars tested：‘2137＇, ‘Akron＇,‘Ankor’,‘ Halt’ ,‘ Jagger’ ,‘ Prairie Red’ , ‘Stanton＇,‘TAM 107＇,‘TAM 110＇,‘Trego＇, ‘ Yuma＇, and ‘Yumar＇. Fewer biotype I apterae responded （settled and fed） in the combination containing more resistant （Dn4- and Dny-expressing） cultivars, compared to the combinations that had fewer. The reverse was true for biotype 2 apterae; more aphids responded in the combination containing the largest number of Dn4 expressing cultivars. Differential colonization of cultivars was observed in only one combination, in which biotype 2 apterae colonized Akron and Yumar in larger numbers than they did Stanton and Yuma. A separate experiment confirmed that, 48 hours after infestation, more biotype 2 apterae abandoned plants of Yuma than plants of Yumar. This differential response was likely due to genetic differences between the two ＇ near isogenic＇ lines that include the lack of Dn4 expression in Yuma. Choice tests with alatae did not result in differential rates of cultivar colonization by either biotype in any combination tested. These results suggest that young wheat plants appear to lack any meaningful antixenosis toward D. noxia, even though the aphids appear to perceive, and sometimes respond to, certain differences in cultivar suitability.     

The influence of host and non‐host companion plants on the behaviour of pest insects in field crops

Companion plants grown as ‘trap crops’ or ‘intercrops’ can be used to reduce insect infestations in field crops. The ways in which such reductions are achieved are being described currently using either a chemical approach, based on the ‘push‐pull strategy’, or a biological approach, based on the ‘appropriate/inappropriate landing theory’. The chemical approach suggests that insect numbers are reduced by chemicals from the intercrop ‘repelling’ insects from the main crop, and by chemicals from the trap‐crop ‘attracting’ insects away from the main crop. This approach is based on the assumptions that (1) plants release detectable amounts of volatile chemicals, and (2) insects ‘respond’ while still some distance away from the emitting plant. We discuss whether the above assumptions can be justified using the ‘appropriate/inappropriate landing theory’. Our tenet is that specialist insects respond only to the volatile chemicals released by their host plants and that these are released in such small quantities that, even with a heightened response to such chemicals, specialist insects can only detect them when a few metres from the emitting plant. We can find no robust evidence in the literature that plant chemicals ‘attract’ insects from more than 5 m and believe that ‘trap crops’ function simply as ‘interception barriers’. We can also find no evidence that insects are ‘repelled’ from landing on non‐host plants. Instead, we believe that ‘intercrops’ disrupt host‐plant finding by providing insects with a choice of host (appropriate) and non‐host (inappropriate) plant leaves on which to land, as our research has shown that, for intercropping to be effective, insects must land on the non‐host plants. Work is needed to determine whether non‐host plants are repellent (chemical approach) or ‘non‐stimulating’ (biological approach) to insects.     

不同光敏色素分子的特异感光性

简要综述两种光敏色素（ＰｈｙＡ、ＰｈｙＢ） 的分子特性、感光性及作用模式等方面的研究进展。光敏色素是一种调节植物中许多光反应的色素蛋白复合体。不同光敏色素分子具有特异的感光性。ＰｈｙＡ负责‘甚低辐照反应’和远红光‘高辐照反应’; 而ＰｈｙＢ则调节‘低辐照反应’及红光‘高辐照反应’。另外, 讨论了ＰｈｙＡ和ＰｈｙＢ在光周期感受中的作用。     

Allometry of Salsola collina in response to soil nutrients,water supply and population density

The allometry of greenhouse‐grown Salsola collina Pall. in response to variation in soil nutrient content, water supply and population density has been compared. The results showed that the biomass allocation was size‐dependent. Root, stem, leaf and reproductive allocation showed a ‘true’ plasticity in response to soil nutrient variation. At low soil nutrient content, plants tended to allocate more biomass to the development of reproductive organs than to stem and leaf, but root allocation was consistent due to a tradeoff between the effects of plant size and soil nutrient content. The plasticity of stem allocation and reproductive effort was ‘true’, while the plasticity of root allocation was ‘apparent’, but there was no plasticity for leaf allocation in response to soil water variation. At lower soil water content, plants tended to allocate more biomass to the stem than to development of reproductive organs. With the exception of ‘apparent’ plasticity of root allocation, no plasticity was detected in biomass allocation when population density was varied.     

Digest: Plant‐pathogen coevolution extends to shifts in plant breeding systems*

Plants and their pathogens are in a coevolutionary arms race. Some pathogens, such as anther smuts, use their host plants’ pollinators for spore dispersal. In the plant Dianthus pavonius, gynodioecy (having female and hermaphroditic plants) has evolved to reduce flowering duration and therefore limit exposure to anther smut pathogens. Bruns et al. (2019) show that this shift in breeding system has evolved as a disease escape mechanism.