Fernanda G. González

Autophagy regulated by day length determines the number of fertile florets in wheat

SUMMARY The wheat spikelet meristem differentiates into up to 12 floret primordia, but many of them fail to reach the fertile floret stage at anthesis. We combined microarray, biochemical and anatomical studies to investigate floret development in wheat plants grown in the field under short or long days (short days extended with low-fluence light) after all the spikelets had already differentiated. Long days accelerated spike and floret development and greening, and the expression of genes involved in photosynthesis, photoprotection and carbohydrate metabolism. These changes started while the spike was in the light-depleted environment created by the surrounding leaf sheaths. Cell division ceased in the tissues of distal florets, which interrupted their normal developmental progression and initiated autophagy, thus decreasing the number of fertile florets at anthesis. A massive decrease in the expression of genes involved in cell proliferation, a decrease in soluble carbohydrate levels, and an increase in the expression of genes involved in programmed cell death accompanied anatomical signs of cell death, and these effects were stronger under long days. We propose a model in which developmentally generated sugar starvation triggers floret autophagy, and long days intensify these processes due to the increased carbohydrate consumption caused by the accelerated plant development.

Wheat floret survival as related to pre-anthesis spike growth

Further improvements to wheat yield potential will be essential to meet future food demand. As yield is related to the number of fertile florets and grains, an understanding of the basis of their generation is instrumental to raising yield. Based on (i) a strong positive association between the number of fertile florets or grains and spike dry weight at anthesis; and (ii) the finding that floret death occurs when spikes grow at maximum rate, it was always assumed that floret survival depends on the growth of the spike. However, this assumption was recently questioned, suggesting that assimilates diverted to the spike do not determine the number of florets and grains and that the onset of floret death may instead be a developmental process that is not associated with spike growth. In this study, the relationships between the fate of floret primordia and spike growth from six independent experiments that included different growing conditions (greenhouse/field experiments, growing seasons, photoperiod/shading treatments during the floret primordia phase) and diverse cultivar types (winter/spring, semi-dwarf/standard-height, photoperiod sensitive/insensitive) were re-analysed together. Onset of floret death was associated with the beginning of spike growth at the maximum rate in c. 80% of the cases analysed; and the rate of floret death (the main determinant of floret survival) showed a negative quantitative relationship with spike weight at anthesis. As floret death and survival were shown to be linked to pre-anthesis spike growth, the strategy of focusing on traits associated with pre-anthesis spike growth when breeding to increase wheat yield potential further is valuable.

Photoperiod Sensitivity during Stem Elongation as an Avenue to Raise Potential Yield in Wheat

Worldwide wheat yields have been only slightly, and non-significantly, increasing during the 90’s, suggesting that they may be levelling off. Considering that there is consensus that large new growing areas will be not introduced and management improvements will be increasingly harder to obtain, genetic improvement would play a more important role to keep rising wheat yields in the future than in the past. In this scenario, the use of physiological bases as a complementary tool to identifying alternative ways for breeding seems to be crucial for breaking the apparent barriers in wheat yields. In this presentation, we attempted to envisage, from published and recent unpublished evidences from our lab, using studies carried out under both controlled and field conditions, some opportunities to manipulate the rate of crop development during the late reproductive phase. This phase has been recognised as critical in terms of yield generation, and the idea of manipulating its response to photoperiod as a tool for increasing yield potential in wheat is reviewed.

Pre-anthesis development and number of fertile florets in wheat as affected by photoperiod sensitivity genes Ppd-D1 and Ppd-B1

Lengthening the late reproductive phase (LRP) of stem elongation in wheat (Triticum aestivumL.), by changing its photoperiod sensitivity independently of the preceding phases, would improve the yield potential through increasing spike weight and the number of fertile florets at anthesis. This paper presents results of a two-year field experiment designed to determine the impact of Ppd-D1and Ppd-B1on (i) the duration of three pre-anthesis developmental phases, and (ii) spike weight and the number of fertile florets at anthesis under two photoperiods during the LRP (natural and an extension of six hours over that). Near isogenic lines of Mercia and single chromosome recombinant lines of Cappelle Desprez were used. Under natural photoperiod, Ppd-D1hastened time to anthesis ca. 500∘C d in both backgrounds by reducing each of the three pre-anthesis phases. Ppd-B1hastened the time to anthesis under natural photoperiod by 178∘C d, mainly by reducing the early reproductive phase. The response to photoperiod of the LRP under extended daylength depended on the Ppdlocus present: Ppd-D1was insensitive while Ppd-B1and the recessive controls were sensitive. For all lines, photoperiod treatments and years, the number of fertile florets was associated with spike dry weight at anthesis (R2≅ 80%, p< 0.01) which, in turn, was positively related to the intercepted radiation accumulated during the LRP (R2 45%, p< 0.05). Changing the duration of the LRP through extended photoperiod or through Ppd-D1produced similar results in both backgrounds and years. Thus, altering the duration of the LRP by manipulating photoperiod sensitivity may be an alternative to changing the fertile floret number in wheat. Nevertheless, as no particular allele was responsible for the photoperiod sensitivity only during the LRP, new alleles should be studied to identify the control of photoperiod sensitivity of individual phases to fine-tune the pre-anthesis wheat development.

Vernalization and photoperiod responses in wheat pre-flowering reproductive phases

Abstract It has been established that photoperiod influences the rate of wheat development well beyond the end of the vegetative phase. Conversely, vernalization effects are still assumed to be mainly during the vegetative phase. The aim of the study was to assess the effects of different combinations of vernalization and photoperiods on the developmental characteristics of wheat pre-flowering reproductive phases under field conditions. For this purpose, three high yielding cultivars (ProINTA Puntal, ProINTA Super and Klein Pegaso) were subjected to two vernalization treatments (unvernalized (V0) and vernalized during 56 days in a cool chamber (V56)) and four photoperiod regimes. The photoperiod treatments consisted of the natural photoperiod of the season (NP+0) and daylength extensions over that of 2 (NP+2), 4 (NP+4), and 6 (NP+6) hours. Photoperiod influenced the duration of early (collar—first node detectable) and late (first node detectable—heading) pre-flowering reproductive phases, reducing their durations as photoperiod increased. When cultivars with strong vernalization response (ProINTA Puntal and ProINTA Super) were not vernalized, duration of the early and late pre-flowering reproductive phases increased and the rate of spikelet initiation decreased. Thus, when vernalization requirements were not satisfied, the spikelet initiation rate was even slower than the leaf initiation rate (both expressed in a thermal time base). Results showed significant interactions between photoperiod and vernalization (vegetative phase), or clear, though non-significant, trends to interactive effects (late pre-flowering reproductive phase). Thus, the length of vegetative and late pre-flowering reproductive phases changed in response to photoperiod depending on the level of satisfaction of the vernalization requirements. The relationship between the cumulative number of leaves on the main stem and thermal time fitted a bi-linear model whenever the final leaf number (LNf) was large, e.g. when photoperiod was not extended or vernalization requirements not satisfied. Thus, responses during the pre-flowering reproductive phases were accompanied by an increased phyllochron for leaves emerging after the first 7–8 leaves. This study demonstrated with field grown plants that the length of the late pre-flowering reproductive phase in wheat is sensitive to photoperiod and that the response to this environmental factor could be changed by the level of satisfaction of the vernalization requirements.

Floret development and survival in wheat plants exposed to contrasting photoperiod and radiation environments during stem elongation

Wheat breeding has improved yield potential increasing floret survival through higher dry matter partitioning to the spikes during the stem elongation phase (from terminal spikelet initiation to anthesis). We studied survival of floret primodia in different spikelet positions along the spike in relation to dynamics of spike growth, when dynamics of dry matter partitioning to the spike was altered by photoperiod and shading treatments applied during the stem elongation phase. The cultivar Buck Manantial was exposed to (1) NP+0 un-shaded (natural photoperiod and incoming radiation of the growing season), (2) NP+0 shaded (natural photoperiod but only 33% of the incoming radiation), and (3) NP+6 un-shaded (natural photoperiod extended 6 h and natural incoming radiation). Floret survival increased, depending on spikelet position, 1.1-2.5 fold under un-shaded v. shaded treatments (both under NP+0), and 1.3-1.8 fold under NP+0 v. NP+6 treatments (both un-shaded), without any impact of treatments on the total number of initiated floret primordia. The fate of the floret primordia and its final stage at anthesis were associated with duration of floret development within the stem elongation phase (R2 = 82%, P<0.0001). Florets may be classified into three groups: (i) those that were fertile at anthesis under all treatments (mostly the two florets F1 and F2, proximal to the rachis within the spikelet), (ii) those that reached different stages at anthesis, depending on treatment, and that contributed differentially to the number of fertile florets at anthesis (mostly the florets F3, F4 and F5, positioned in the middle of the spikelet), and (iii) those that did not contribute to the number of fertile florets under any treatment (mostly the florets ≥ F6). Degeneration of florets in group (ii) was associated with spike growth at maximum rate, explaining the strong relationship observed between spike dry weight at anthesis and number of fertile florets. However, degeneration of florets in group (iii) seemed to occur before spike growth at maximum rate. Survival of florets positioned in the middle of the spikelets could be improved by increasing spike growth through manipulation of photoperiod sensitivity during stem elongation.

Photoperiod during stem elongation in wheat: is its impact on fertile floret and grain number determination similar to that of radiation?

Increasing duration of stem elongation by exposure to short photoperiod would result in higher spike dry weight at anthesis, which is positively associated with the number of fertile florets and grains in wheat. However, it is not easy to determine whether photoperiod effects on fertile florets and grains are only mediated by assimilate supply to the growing spike when spike weight variation is attained only with photoperiod treatments. The aim of this study was to determine whether photoperiod effects on number of fertile florets and grains may be direct, that is, not mediated by assimilate supply, by comparing the magnitude of photoperiod effects with those of shading the canopy. Spike dry weight at anthesis was changed through the factorial combination of different photoperiod (natural and 6 h extended photoperiod) and shading (un-shaded and 67 ± 3% shaded) treatments during stem elongation of Buck Manantial, a cultivar known for its photoperiod sensitivity in this phase. Both treatments modified spike dry weight at anthesis and the number of fertile florets and grains, independently. When duration of stem elongation was lengthened by exposure to natural photoperiod and when incident radiation was high, spike dry weight at anthesis increased by 33% (NP+0 v. NP+6) and 27% (un-shaded v. shaded), respectively. The number of fertile florets increased similarly to spike dry weight (34% NP+0 v. NP+6 and 28% un-shaded v. shaded) resulting in higher number of grains. Most photoperiod effects on the number of fertile florets and, consequently, on the number of grains, were mediated by assimilate supply to the growing spike as the same relationship between the number of fertile florets and spike dry weight at anthesis was observed for photoperiod and shading treatments (R2 = 0.99, P<0.05).

Strategic crossing of biomass and harvest index—source and sink—achieves genetic gains in wheat

To accelerate genetic gains in breeding, physiological trait (PT) characterization of candidate parents can help make more strategic crosses, increasing the probability of accumulating favorable alleles compared to crossing relatively uncharacterized lines. In this study, crosses were designed to complement “source” with “sink” traits, where at least one parent was selected for favorable expression of biomass and/or radiation use efficiency—source—and the other for sink-related traits like harvest-index, kernel weight and grains per spike. Female parents were selected from among genetic resources—including landraces and products of wide-crossing (i.e. synthetic wheat)—that had been evaluated in Mexico at high yield potential or under heat stress, while elite lines were used as males. Progeny of crosses were advanced to the F4 generation within Mexico, and F4-derived F5 and F6 generations were yield tested to populate four international nurseries, targeted to high yield environments (2nd and 3rd WYCYT) for yield potential, and heat stressed environments (2nd and 4th SATYN) for climate resilience, respectively. Each nursery was grown as multi-location yield trials. Genetic gains were achieved in both temperate and hot environments, with most new PT-derived lines expressing superior yield and biomass compared to local checks at almost all international sites. Furthermore, the tendency across all four nurseries indicated either the superiority of the best new PT lines compared with the CIMMYT elite checks, or the superiority of all new PT lines as a group compared with all checks, and in some cases, both. Results support—in a realistic breeding context—the hypothesis that yield and radiation use efficiency can be increased by improving source:sink balance, and validate the feasibility of incorporating exotic germplasm into mainstream breeding efforts to accelerate genetic gains for yield potential and climate resilience.

Wheat pre-anthesis development as affected by photoperiod sensitivity genes (Ppd-1) under contrasting photoperiods

Fine tuning wheat phenology is of paramount importance for adaptation. A better understanding of how genetic constitution modulates the developmental responses during pre-anthesis phases would help to maintain or even increase yield potential as temperature increases due to climate change. The photoperiod-sensitive cultivar Paragon, and four near isogenic lines with different combinations of insensitivity alleles (Ppd-A1a, Ppd-B1a, Ppd-D1a or their triple stack) were evaluated under short (12h) and long (16h) photoperiods. Insensitivity alleles decreased time to anthesis and duration of the three pre-anthesis phases (vegetative, early reproductive and late reproductive), following the Ppd-D1a > Ppd-A1a > Ppd-B1a ranking of strength. Stacking them intensified the insensitivity, but had no additive effect over that of Ppd-D1a. The late reproductive phase was the most responsive, even exhibiting a qualitative response. Leaf plastochron was not affected but spikelet plastochron increased according to Ppd-1a ranking of strength. Earlier anthesis resulted from less leaves differentiated and a fine tuning effect of accelerated rate of leaf appearance. None of the alleles affected development exclusively during any particular pre-anthesis phase, which would be ideal for tailoring time to anthesis with specific partitioning of developmental time into particular phases. Other allelic variants should be further tested to this purpose.

Correction to: Strategic crossing of biomass and harvest index—source and sink—achieves genetic gains in wheat

The original article was corrected. Author Muhammad Kundi should instead read: Muhammad Sohail.