ITEMS FROM THE RUSSIAN FEDERATION


ALL-RUSSIAN INSTITUTE OF AGRICULTURAL BIOTECHNOLOGY -ARRIAB

Timiryazevskaya ul. 42, Moscow, 127550, Russia.

The problem of female fertility and some possibilities to improve Triticum aestivum L. x T. timopheevii Zhuk. hybrids at the first backcross.

V.F. Kozlovskaya and M.M. Starostenkova.

Introgressive hybrids are known to be characterized by a self-sterile first generation, and progeny usually is obtained by backcrossing. This undoubtedly important stage is the least studied step in the introgression of alien genetic material into cultivated wheat, because of parental incompatibility and low F1 hybrid production. In our experiments, we have overcome the incompatibility between T. aestivum and T. timopheevii. The main objectives of this study were to analyze variation in the first backcross, determine factors to correct them, and ascertain the reasons for infertility in F1 `T. aestivum x T. timopheevii' hybrids using cytoembryological techniques.

Six bread wheat cultivars, derived from different ecological areas, were used as maternal and recurrent parents. Triticum timopheevii var. viticulosum, K-47793, was used as a pollinator. Backcrosses were by conventional methods (a one-time pollination at the beginning of flowering) and also using an optimum pollinating regime. The optimum pollinating time was determined by comparing the BC1 effectiveness after twofold pollinating (on the first (I) and third (III) days and on the second (II) and fourth (IV) days from the hybrids' anthesis) concurrent with daily pollinating (IV) in experimental conditions. The same set of hybrids was backcrossed using conventional (4 seasons) and optimum pollinating (3 seasons) regimes during 4 and 3 seasons, respectively. An analysis of variance was made on the experimental data.

After conventional backcrossing, an analysis of variation showed that the prevalent contribution to phenotypic variability belonged to environmental factors. The average proportion of the environmental contribution exceeded that of `genotype' and `genotype x environment' interaction by 3.7 times. Consequently, an increase in BC1F1 seed set after conventional backcrossing was due to positive modifications. We hypothesize that the environment affected the results through anthesis dynamics. Optimizing a pollinating regime according to flowering dynamics could lead to a reduction in environmental effects and to an increase in seed set. Based on the high seed set, labor, time, and material consumption, we established that pollinating regime I, III was optimum. This regime ensured a 1.8-times increase (from 3.3 to 6.0 %) of the results on average of the whole hybrid seed set. This suitable pollination regime was used when six F1 hybrids were backcrossed for three seasons. A 3.7-times increase of `genotype' contribution and more than a 5-times (from 37.0 to 7 0 %) reduction of the `environmental' component were found through statistical analysis. Hence, anthesis dynamics appeared to be environmentally dependent and impeded the reliable estimation of the genotype's role in variation of the studied trait.

A cytoembryological analysis of six F1 `T. aestivum x T. timopheevii' hybrids indicated that the specific disturbance was the lack of an embryo sac. The hybrid frequency was about 79.9 and varied from 70.4 to 97.5, according to genotype. This abnormality in hybrid genotypes could be due to the incapability of the megaspore egg-cell to differentiate or to cell lysis or breaking mechanisms prior to meiosis. In any case, these irregularities took place at the sporophyte stage, and sterility is of a diplontic nature. However, we did not notice this type of abnormality during a mass analysis of PMCs in meiosis of the present hybrids. Probably, this is evidence of independent genetic control of male and female generative sphere development. Available embryo sacs underwent destructive changes to different extents (5.5 % of embryo sacs) and ranged from slightly to extremely visible: nuclei appeared as dark, formless spots on a structureless cytoplasm background. Furthermore, 5.9 % of the embryo sacs lacked antipodes and were referred to as structural failures. Normal female gametophytes were observed with an average frequency of 4.8 % with a limit of variation of 0.8-11.7 %, depending on genotype.

Consequently, the efficiency of introgression at the most difficult stageóthe first backcrossócould be increased greatly by obtaining a set of hybrids to indirectly estimate the level of female fertility, (based on the results of the optimum (I, III) regime of pollination) or directly according to the frequency of embryo sac occurrence in the ovule. The result will be to provide the greatest numbers of F1 seeds for breeding and research with the use of the most suitable genotypes.

Personnel.

This year Dr. Sci. (Biol.) Vera Kozlovskaya continued the research on introgressive hybridization of wheat species. Marina Starostencova is a second year postgraduate student.


THE MOSCOW PEOPLE'S FRIENDSHIP UNIVERSITY

Ul. Efremova, I8, kv. 7, Moscow, 119048, Russia.

Reaction of wheat to light and length of the vegetation period.

Alexander Federov.

The adaptation of plants to environmental conditions, resistance to pathogens and pests, and the amount and quality of harvest are determined to a great extent by the length of the vegetative period. What causes different lengths of the vegetation period are not yet fully known. Although the so-called theory of the stage development of plants (Lysenko, 1936) has been invalidated, publications still appear where authors support this theory.

According to the theory of stage development of plants, the type of development (winter or spring) and length of the vegetation period (early or late ripening) are determined by vernalization, particularly by its duration and conditions (Lysenko, 1936). This corresponds, to a certain extent, to data from the 1930s and 1950s, when it was discovered that winter wheat planted in the spring from vernalized seeds (germinated seeds kept at 0-3_C for 40-60 days) yielded like a spring-planted spring wheat. There are experimental data on the vernalization of spring wheat, but it was believed to be of short duration (5-15 days/year) and required higher temperatures (5-15_C) (Lysenko, 1936).

The basis of this theory disagrees with the published data:

1. No reaction to vernalization occurs under normal growing conditions in many spring cereals (Gypalo and Skripchinsky 1971).

2. Winter varieties will flower without vernalization when grown under continual, intensive light (Fedorov, 1976, 1989) or when grown first under short days for a certain period followed by growth in long days (Rasunov 1961; Fedorov 1996).

3. A vernalization requirement is lacking under field conditions in cereals and, in many forage grasses, all types of plant development are lacking during the spring-summer period. Spring and alternative plants flower and bear fruits normally without vernalization. Winter-sown alternative and winter plants vernalize in autumn, but do not react to vernalization in spring and summer.

4. The alternative and winter wheats (originating from the same geographical region) as a rule, have identical vernalization requirements (length and conditions). In the field, their vernalization begins and ends at approximately the same time (Fedorov 1976, 1989).

The aims of this work were to define the parameters that influence ontogenesis and to determine the type of plant development and the length of the vegetation period. Wheat varieties (predominantly regionalized) differing in type of development (winter, alternative, and spring) were crossed. The hybrids and parental varieties were planted as nonvernalized and vernalized seeds in the field and in the greenhouse under controlled conditions at various day lengths: natural, 12-hour, and continuous illumination. In the greenhouse, lights equipped with DRI 2000-6 metal-halogen lamps and reflecting mirrors were used at the different light regimes. Besides the standard phenological observations, differentiation of the shoot apex was monitored.

Our results indicate that the length of the period from germination to ear formation in F1 hybrids (winter wheat `Lutescens 329' x spring wheat `Lutescens 62') at a natural day length was 6 days shorter than that of the parental spring variety. Under short-day conditions, the difference was 30 days. The reaction of F1 hybrids to vernalization was more pronounced than that of the parental spring variety, especially under short-day conditions. Vernalization accelerated the development of the spring variety by 9 days at a 12-hour illumination, compared to the natural day length, and that of the F1 plants by 20 days.

The following winter wheat cultivars were used: Lutescens 329, with the highest level of winter- and frost-hardiness; Mironovskya 808, of medium winter hardiness; Bankuti 1201, the least winter- and frost-hardy. The alternative wheats used were Czech, with the highest reaction to photoperiod and of medium winterhardiness; Surkhak 5688, a central Asian cultivar; and a Bulgarian cultivar, 109, with the least photoperiodic response and low frost hardiness. The spring wheat Lutescens 62, with a low photoperiodic reaction and poor frost hardiness, was used.

The data of Table 1 suggest that hybrid F1 plants resulting from crossing varieties of different developmental types (spring, winter, and alternatives) are distinguished from each other, and from the parental forms, by their reaction to light and their response to vernalization.

In particular, the early generations from crosses between the alternative and spring cultivars and the winter and spring cultivars under short-day conditions significantly lag behind the spring varieties in development. First generation hybrids from crosses of a winter cultivar with an alternative have a greater lag in development under short days than the hybrid plants of the aforementioned combinations.

The F1 from crosses between winter and alternative cultivars are delayed during short, and also long (natural summer), days. After spring sowing, the F1 hybrids of crosses between winter and alternative cultivars have a long tillering stage similar to that of a winter wheat. But quite unlike a winter wheat, these cultivars form ears at the end of summer, the date depending on both the alternative and the winter cultivars. When the same alternative cultivar (Czech alternative) was crossed with different winter wheats, the duration of the vegetative period and the expression of a photoperiodic response were greater when the degree of winterhardiness was greater, i.e., in those wheats from the farthest north. In particular, the F1 of crosses between the Czech alternative and Lutescens 329 formed ears later and lagged in development more with short days than the F1 of crosses with Bankuti 1201. The degree of the photoperiodic response (a greater lag of development with short days) in the F1 was higher than in the parent alternative wheat, because of the influence of the winter parent.

Table 1. The influence of day length and vernalization on the development of the F1 from crosses between

spring (S), winter (W), and alternative (A) wheat cultivars (sown 30 April).

_________________________________________________________________________________________

Time from

emergence to

Light differentiation Heading

Cross Seed sown treatment of shoot (days) date

_________________________________________________________________________________________

(W x S) F1 Lutescens 329 x Lutescens 62

unvernalized natural 13 30.06

vernalized natural 13 5.07

unvernalized 12 h 29 24.08

vernalized 12 h 18 4.08

(A x S) F1 Czech Alternative x Lutescens 62

unvernalized natural 16 30.06

vernalized natural 14 27.06

unvernalized 12 h 27 23.08

vernalized 12 h 18 5.08

(W x A) F1 Lutescens 329 x Czech Alternative

unvernalized natural 56 20.08

vernalized natural 18 13.07

unvernalized 12 h 86 did not head

vernalized 12 h 30 did not head

(W x A) F1 Bankuti 1201 x Czech Alternative

unvernalized natural 41 1.08

unvernalized 12 h 69 did not head

(W x A) F1 Lutescens 329 x Surkhak 5688

unvernalized natural 39 29.07

unvernalized 12 h 65 did not head

(A) Czech Alternative

unvernalized natural 25 12.07

vernalized natural 14 2.08

unvernalized 12 h 52 did not head

vernalized 12 h 23 16.08

(W) Lutescens 329*

unvernalized natural 119 20.08

vernalized natural 15 13.07

unvernalized 12 h ó did not head

vernalized 12 h 29 did not head

(S) Lutescens 62

unvernalized natural 12 24.06

vernalized natural 12 24.06

unvernalized 12 h 23 26.07

vernalized 12 h 18 20.07

_________________________________________________________________________________________

* Under intensive, continuous light conditions (50-70 klx) and headed on 22 August).

Therefore, an increase in the degree of response to photoperiod in the F1 is due to the winter cultivar. This discovery also is supported by the fact that, unlike the alternative variety, this F1 lags behind in development under long-day conditions, but not as much as the winter variety.

These data suggest that plants of different developmental types, particularly alternative and winter wheats, are distinguished by their reaction to light. The delayed development under short-day conditions in alternative wheats and under both long- and short-day conditions in winter wheats is basically the same phenomenon. Both the reaction of unvernalized alternative wheats to photoperiod (lag of development under short-day conditions) and the winterhardiness of winter wheats (lag of development under long- and short-day conditions) are adaptive properties resulting from the development of winterhardiness in the autumn.

Plant response to vernalization is determined by their reaction to light. The more cultivars and F1 hybrids react to vernalization, the longer the delay in development under definite light conditions. Under long-day conditions, the F1s of the crosses between `spring and alternative' and `spring and winter' wheats show very low, if any, delay in development, with no response to vernalization. The F1s of crosses between `winter and alternative' wheats show a considerably longer delay of development under long-day conditions and, accordingly, acceleration of development after vernalized seed are sown. Under short-day conditions, plants of all hybrid combinations respond to vernalization more effectively than under long days. The lag of development under short days is longer. In their reaction to light and vernalization, the F1s are intermediate between the original parent lines, but closer to that parent with the least degree of winter hardiness.

In the duration and pattern of vernalization, the F1s of the crosses between a winter cultivar and an alternative show almost no difference from the parent varieties. For example, the F1s of crosses of the winter wheat Mironovskya 808 with the Czech alternative has a 45-day vernalization, equal to that of the parents. However, the F1s and parents differ markedly in development type and vegetative period, mainly because of their different reaction to light conditions related to response to, but not the duration and temperature conditions of, vernalization. In our studies conducted over many years, segregation in F2 hybrids from `spring x winter', `alternative x winter', and `alternative x spring' wheats was always complicated in terms of the type of development, length of the vegetative period, and winterhardiness. Among the hybrid plants, there usually was a complete transition from one parental variety to another (Table 2).

Hybrid F2 plants from the crosses `Lutescens 329 x Lutescens 62' and `Lutescens 329 x Czech alternative' were divided into nine and eight classes, respectively, according to the duration of the vegetative period or, more precisely, the period from the development of full sprouts to ear formation (Table 2). A daily count of the number of plants that formed ears among the F2 hybrids from crosses between `winter x spring' and `winter x alternative' wheats allowed us to distinguish 20 groups of plants that differed in terms of the duration of the period from the development of full sprouts to ear formation. In the latter cross, there were no spring plants, only alternative, semiwinter, and winter plants.

All F2 hybrids from `alternative x spring' crosses formed ears, although not simultaneously. There were no winter plants among these hybrids. Most of the F2 plants were intermediate between the parents in terms of the duration of the vegetative period. In one experiment, the duration of the vegetative period was the same as in the spring wheat in 10 % of the plants and the same as the Czech alternative in 6 % of the plants, whereas 84 % of the plants had intermediate duration of vegetative period. Some winter wheats from Bulgaria, Hungary, Serbia, and other relatively southern regions, (including Southern Russia) planted in the Moscow District in spring all formed ears at the end of summer-autumn. In experiments conducted over many years, we found this to be true of the varieties

Bankuti 1201, Bezostaya I, Aurora, Kooperatorka, and Sava. All F2 hybrids from crosses of these cultivars with spring types usually formed ears. They are divided into 7-12 classes (10-day intervals). The vast majority of

Table 2. Distribution of the F2 hybrids by classes (10-day interval) according to the length of the period from development of full sprouts to ear formation.

___________________________________________________________________________________________

Number of plants in class

Number

__________________________________________ Number

of 40- 50- 60- 70- 80- 90- 100- 110- winter

Cross plants 49 59 69 79 89 99 111 119 plants

___________________________________________________________________________________________

Lutescens 239 x Lutescens 62 366 85 152 35 18 12 12 8 10 34

Lutescens 329 x Czech alternative 367 0 10 152 31 13 10 11 12 128

Czech alternative x Lutescens 62 320 221 99 0 0 0 0 0 0 0

___________________________________________________________________________________________

hybrid plants were intermediate between the parents in terms of the duration of the period from the development of full sprouts to ear formation. Usually, the duration of this period in 50 to 60 % of the plants was longer than that of the spring wheat by 5-15 days, in 4 % of the plants equaled that of the spring variety, and in 8 % equaled that of the winter cultivar.

Winter wheats, ryes, Triticales, and barleys formed ears at the same time (Fedorov 1959, 1960, 1989) at relatively high temperatures under conditions of continuous intense illumination (approximately 50 klk) without previous vernalization. This also was true of F2 hybrids from crosses of `winter x spring' types. The duration of the vegetative period varied. In F2 hybrids of the cross `Lutescens 329 x Lutescens 62', 18 groups of plants were distinguished that differed in the duration of the period from the development of full sprouts to ear formation. This period lasted 3 days in Lutescens 62 and 116 days in Lutescens 329. Among the F2 hybrids, there was almost complete transition from one parental variety to the other.

Two photoperiodic responses occur in plants: a strongly expressed reaction in nonvernalized plants and a weakly expressed reaction in vernalized plants. The former determines the duration of the vegetative period in plants of spring planting, and the second determines the same in plants of autumn planting.

Type of development (winter, alternative, or spring) and duration of the vegetative period are determined by the reaction of the plants to light during the initial period (tillering), rather than by vernalization, as was suggested by Lysenko (1936) and Pugsley (1971). Unlike spring cereals, winter cereals (winter and alternative) were delayed during tillering (winter wheat under both long- and short-day conditions and alternative wheat under short-day conditions), which enhanced their winter hardiness. Spring wheats reacted weakly to short-day conditions.

The difference in length of the vegetative period of wheats and other plants (as type of development, resistance against unfavorable environmental conditions) are determined to a great extent by delay of the transition of the shoot apex from the vegetative to the generative phase. The vegetative phase is the best time for the adaptation of plants to environmental conditions; after transition to the generative phase, plants lose the ability to adapt.

References.

Fedorov AE. 1976. On photoperiodism, wintering and vernalization in wheat. Cereal Res Commun 4(4):419.

Fedorov AE. 1989. Fodder Plants. Moscow Nauka. 160 pp.

Fedorov AE. 1989. Physiological-genetical basis for the type of plant development and length of the vegetation period in wheat. Cereal Res Commun 7(2):121.

Gupalo PI and Skripchinskii VV. 1971. Physiology of Plant Development. Moscow Kolos 224 pp.

Lysenko TD. 1936. Theoretical Foundations of Vernalization. Moscow Selckhozgiz 94 pp.

Pugsley AT. 1971. Genetical analysis of the spring-winter habit of growth. Austral J Agric Res 22(1):21.

Razumov VI. 1961. Environmental and Plant Development. Moscow Selckhozgiz 368 pp.


N.I. VAVILOV INSTITUTE OF GENERAL GENETICS

Gubkin st.3, 117809 Moscow, Russia, and the Agricultural Research Institute of the Non-Chernozem Zone, Nemchinovka, 143013 Moscow region, Russia.