Phenology and dry mass production of Urochloa plantaginea and Urochloa platyphylla submitted to different water quantities in the soil

The genetic variability and genetic versatility of plants belonging to Poaceae provide morphophysiological responses that allow these individuals to adapt to environmental changes, especially in relation to soil moisture. Urochloa plantaginea and U. platyphylla are grasses typical of dry environment, also found as weeds in rice fields, where there are high amounts of water in the soil. The objective of this work is to analyze the development of these two species in different environments, subjecting them to three different amounts of water in the soil. Morphological parameters were analyzed in order to verify and quantify which suffered alteration with respect to water variation. Caryopsis of the two species were collected in a commercial irrigated rice crop. Seeds were sown in pots containing a substrate-sand system, and housed in a greenhouse, where during the experiment the control of different moisture contents (shallow water table, 100% of field capacity and 50% of field capacity) took place. Weekly evaluations were carried out according to the parameters analyzed. The following parameters were evaluated: duration of the vegetative and reproductive life cycle, number of the inflorescences per plant, number of branches per inflorescence, number of spikelets per branch, number of seeds per plant, and dry mass production. The U. plantaginea cycle had shorter duration under the shallow water table, while for U. platyphylla the shorter duration was under the condition 50% of field capacity. Both species produced higher dry mass of shoots under the condition of 100% of field capacity. The main responses observed for both species, due to the flooding, were the change of the cycle, reduced dry matter production, and reduced seed production.

Regrowth age modifies the leaf anatomy of Brachiaria genotypes / A idade de regeneração modifica a anatomia foliar dos genótipos de Brachiaria

Changes in leaf anatomy were evaluated in genotypes of Brachiaria brizantha, Brachiaria decumbens, Brachiaria ruziziensis and three Brachiaria ruziziensis clones through tissue proportion in internerval and midrib regions at three regrowth ages. Plants were grown and cutting was performed after 60 days. Further, leaves were sampled at 8, 15 and 29 regrowth days and processed with usual plant microtechnique. The internerval region showed higher parenchyma percentage at 15 days for Clone 95 and similar values at 15 and 29 days for Clone 1. The proportion of vascular bundles was lower after 15 days in Clones 1 and 95 and 29 days in B. brizantha. In the midrib, the parenchyma proportion was higher at 29 days in B. brizantha and lower at 15 days in B. ruziziensis. The proportion of vascular bundles was higher at 8 days in B. decumbens, B. brizantha and Clone 1, and lower at 29 days for Clones 97 and 95 and at 8 days in B. ruziziensis. Therefore, the regrowth age modifies the percentage of leaf tissues in Brachiaria genotypes, in which the fibers and vascular bundles increase at 29 days and 8-day-old leaves are not fully developed.

A anatomia foliar foi avaliada em genótipos de Brachiaria brizantha, Brachiaria decumbens, Brachiaria ruziziensis e três clones de Brachiaria ruziziensis; quanto à proporção de tecidos foliares, em três idades de rebrota. As plantas foram cultivadas e cortadas após 60 dias. As folhas foram amostradas aos 8, 15, 29 dias de rebrota e analisadas pela metodologia de microtécnica vegetal. Na região internervural, a proporção de parênquima foi maior aos 15 dias para o Clone 95, e aos 15 e 29 dias para o Clone 1. Proporção de feixes vasculares foi menor aos 15 dias para os Clones 1 e 95 e aos 29 dias para genótipo B. brizantha. Na nervura central, a proporção de parênquima foi maior aos 29 dias para B. brizantha e menor, aos 15 dias, para B. ruziziensis. Proporção dos feixes vasculares foi maior aos 8 dias para genótipo B. decumbens, B. brizantha e para o Clone 1, e menor aos 29 dias para os Clones 97 e 95; aos 8 dias, para genótipo B. ruziziensis. Portanto, a idade de rebrota modifica a porcentagem de tecidos foliares em genótipos de Brachiaria; contudo, fibras e feixes vasculares aumentam aos 29 dias; aos 8 dias, as folhas não estão totalmente desenvolvidas.

Chromosome number and microsporogenesis of two accessions of Brachiaria dura Stapf (Poaceae) / Número de cromossomos e microsporogênese de dois acessos de Brachiaria dura Stapf (Poaceae)

The two accessions of B. dura analyzed (DU01 and DU02) are hexaploid (2n = 6x = 54), derived from x = 9. Meiotic abnormalities, such as precocious chromosome migration to the poles, laggards and micronuclei, were recorded in low frequency in both accessions. The few multivalent chromosome association at diakinesis and meiotic stability suggested that hexaploidy probably resulted from chromosome doubling. In DU02, chromosome transfer (cytomixis) among meiocytes, involving part or the entire genome was observed. The implication of these findings for the Brachiaria breeding is discussed.

Os dois acessos de B. dura analisados (DU01 e DU02) são hexaplóides (2n = 6x = 54), derivados de x = 9. Anormalidades meióticas como migração precoce de cromossomos para os polos, cromossomos retardatários e micronúcleos foram observados em baixa frequência em ambos os acessos. A presença de poucas associações cromossômicas em diacinese e a estabilidade meiótica sugere que a hexaploidia provavelmente resultou de duplicação cromossômica. No acesso DU02 observou-se transferência de cromossomos (citomixia) entre meiócitos, envolvendo parte ou todo o genoma. As implicações destes resultados para o melhoramento de Brachiaria são discutidas.

Evaluation of microsporogenesis in an interspecific Brachiaria hybrid (Poaceae) collected in distinct years

Microsporogenesis in an interspecific Brachiaria hybrid, grown in the field under natural environmental conditions in Brazilian savannas, was analyzed in three distinct years of collection. Several types of meiotic abnormalities were recorded during those three years, but varied in type and frequency depending on the year. The average temperature and rainfall 15 days before collection was unusually high in those years. The percentage of abnormal meiocytes recorded was 62% in 2001, 73% in 2004, and 77% in 2005. The abnormalities observed during microsporogenesis compromised pollen viability by generating unbalanced gametes or affecting nucleolus organization. The environmental conditions under which the hybrid was growing could have affected the genetic control of meiosis. More detailed studies, under controlled conditions, are necessary to better understand the effects of environmental factors on Brachiaria microsporogenesis hybrids.

Mitotic behavior in root tips of Brachiaria genotypes with meiotic chromosome elimination during microsporogenesis

Three accessions of Brachiaria brizantha, three of B. humidicola, and two interspecific hybrids between B. ruziziensis and B. brizantha were analyzed with regard to their mitotic behavior in root tips. All these genotypes revealed chromosome elimination or lack of chromosome affinity in previous analyses of microsporogenesis. Analyses of root tips showed a normal mitotic division in all accessions and hybrids, reinforcing the notion that the genetic control of meiosis is totally independent of that of mitosis. The implications of these findings for the Brachiaria breeding program are discussed.

Microsporogenesis in sexual Brachiaria hybrids (Poaceae)

Three sexual interspecific hybrids of Brachiaria (HBGC076, HBGC009, and HBGC014) resulting from crosses between B. ruziziensis (female genitor) and B. decumbens and B. brizantha (male genitors) produced by Embrapa Beef Cattle in the 1980s were cytologically analyzed by conventional methods for meiotic studies. The cytogenetic analysis showed the occurrence of common meiotic abnormalities among them. The most frequent abnormalities were those related to irregular chromosome segregation due to polyploidy. Other abnormalities, such as chromosome stickiness, absence of cytokinesis, irregular cytokinesis, abnormal spindle orientation, and abnormal nucleolus disintegration, were found in the three hybrids, while, chromosome disintegration was detected only in HBGC014. All the abnormalities, except for abnormal nucleolus disintegration, can cause unbalanced gamete formation, leading to pollen sterility. Multivalent chromosome association at diakinesis revealed genome affinity between the two parental species in the hybrids, suggesting some possibility for gene introgression. Presently, the Brachiaria breeding program has the objective of releasing, primarily, apomictic hybrids as new cultivars since they do not segregate but preserve the genetic makeup indefinitely. Besides, they result in homogeneous pastures which are easier to manage. The sexual hybrids, however, are paramount in the breeding program: they work as ‘bridges’ to introgress traits of interest into the apomictic genotypes. The cytogenetic analyses of these three hybrids substantiate their maintenance in the breeding program due to low frequency of meiotic abnormalities, complemented by interesting agronomic traits. They may be used in crosses to generate new cultivars in the future.

Microsporogenesis in Brachiaria dictyoneura (Fig. & De Not.) Stapf (Poaceae: Paniceae)

icrosporogenesis was analyzed in five accessions of Brachiaria dictyoneura presenting x = 6 as the basic chromosome number. All accessions were tetraploid (2n = 4x = 24) with chromosome pairing in bi-, tri-, and quadrivalents. The recorded meiotic abnormalities were those typical of polyploids, including precocious chromosome migration to the poles, laggard chromosomes, and micronucleus formation. The frequency of these abnormalities, however, was lower than those reported for other polyploid accessions previously analyzed for other Brachiaria species. Cell fusion and absence of cytokinesis were also recorded in some accessions, leading to restitutional nucleus formation in some cells. Genetically unbalanced microspores, binucleate, and 2n microspores were found among normal meiotic products as results from these abnormalities. The limitation in using these accessions as pollen donor in interspecific crosses with sexual species with x = 7 or x = 9 in breeding programs is discussed.

Evidence of allopolyploidy in Brachiaria brizantha (Poaceae: Paniceae) through chromosome arrangement at metaphase plate during microsporogenesis

In the hexaploid (2n = 6x = 54) accession B176 of Brachiaria brizantha, one cytological characteristic differentiated it from the other accessions previously analyzed with the same ploidy level. Nearly 40% of meiocytes displayed the chromosome set arranged at two metaphase plates at the poles of the cell, close to the membrane. In these cells, both metaphase plates were arranged in an angle to form a typical tripolar spindle. Therefore, cells did not show normal chromosome segregation at anaphase I. Only nine univalent chromosomes migrated from each plate to the opposite pole with the remainder staying immobile on the plate. As a result of such spindle orientation and chromosome behavior, trinucleate telophases I were recorded. After telophase, cytokinesis eliminated the small nuclei into a microcyte. The second division proceeded normally, with the presence of microcytes in all phases. The origin of such an abnormality was explained on the hexaploid level of the accession which could have resulted by chromosome doubling of a triploid derived from species that did not display the same behavior for spindle organization. The high percentage of meiotic abnormalities recorded in this accession compromises fertility and renders it inadequate for the breeding program.

Abnormal spindle orientation during microsporogenesis in an interspecific Brachiaria ( Gramineae ) hybrid

This paper reports a case of abnormal spindle orientation during microsporogenesis in an interspecific hybrid of the tropical grass Brachiaria. In the affected plant, prophase I was normal. In metaphase I, bivalents were regularly co-oriented but distantly positioned and spread over the equatorial plate. In anaphase I, chromosomes failed to converge into focused poles due to parallel spindle fibers. As a consequence, in telophase I, an elongated nucleus or several micronuclei were observed in each pole. In the second division, the behavior was the same, leading to polyads with several micronuclei. A total of 40 percent of meiotic products were affected. The use of this hybrid in production systems needing good-quality seeds is discussed.

Cytogenetic evidence for genome elimination during microsporogenesis in interspecific hybrid between Brachiaria ruziziensis and B. brizantha (Poaceae)

Microsporogenesis was analyzed in an interspecific hybrid between an artificially tetraploidized sexual accession of Brachiaria ruziziensis (R genome) and a natural apomictic tetraploid accession of B. brizantha (B genome). Chromosomes associated predominantly as bivalents. From this phase to the end of meiosis, chromosomes presented irregular segregation and abnormal arrangement in the metaphase plate. During metaphase I, in 27.8 percent of meiocytes, bivalents were distributed in two metaphase plates. In anaphase I, two distinct and typical bipolar spindles were formed. In 29.7 percent of pollen mother cells, one genome did not divide synchronically, with chromosomes lagging behind or not segregating at all. The second division was very irregular, resulting in polyads. Based on previous results from analysis of a triploid hybrid between these species, where the R genome was eliminated by asynchrony during meiosis, it is suggested that the laggard genome in this hybrid also belongs to B. ruziziensis.

Microsporogenesis in tetraploid accessions of Brachiaria nigropedata (Ficalho & Hiern) Stapf (Gramineae)

The genus Brachiaria (Trin.) Griseb. has achieved considerable importance to cattle production systems, as a result of the good production and adaptation of a few cultivars to poor and acid soils of the Brazilian savannas. Many of its species and accessions are polyploid and apomictic, which limits direct hybridization. To assist the breeding program, cytogenetic characterization has been undertaken on the accessions of Brachiaria collection at the Embrapa Beef Cattle Research Center. In this study, chromosome number and meiotic behavior are reported for the Brachiaria nigropedata (Ficalho & Hiern) Stapf collection. The 20 available accessions are tetraploid (2n = 4x = 36). Chromosomes paired preferentially as bivalents, but quadrivalents were found in high frequencies in some cells. Meiotic behavior was, in general, irregular, and varied among accessions. Most accessions presented more than 20% of abnormal tetrads. The most common meiotic abnormalities were those related to irregular chromosome segregation due to polyploidy, leading to micronuclei formation in the tetrad stage. A low frequency of other meiotic abnormalities such as the absence of cytokinesis, chromosome stickiness, cell fusion, anaphase bridges, and chromosome transfer among microsporocytes were also recorded in some accessions. Limitations of these accessions for use in hybridization programs are discussed.

Symmetric pollen mitosis I and suppression of pollen mitosis II prevent pollen development in Brachiaria jubata (Gramineae)

Microsporogenesis and pollen development were analyzed in a tetraploid (2n = 4x = 36) accession of the forage grass Brachiaria jubata (BRA 007820) from the Embrapa Beef Cattle Brachiaria collection that showed partial male sterility. Microsporocytes and pollen grains were prepared by squashing and staining with 0.5 percent propionic carmine. The meiotic process was typical of polyploids, with precocious chromosome migration to the poles and laggards in both meiosis I and II, resulting in tetrads with micronuclei in some microspores. After callose dissolution, microspores were released into the anther locule and appeared to be normal. Although each microspore initiated its differentiation into a pollen grain, in 11.1 percent of them nucleus polarization was not observed, i.e., pollen mitosis I was symmetric and the typical hemispherical cell plate was not detected. After a central cytokinesis, two equal-sized cells showing equal chromatin condensation and the same nuclear shape and size were formed. Generative cells and vegetative cells could not be distinguished. These cells did not undergo the second pollen mitosis and after completion of pollen wall synthesis each gave rise to a sterile and uninucleate pollen grain. The frequency of abnormal pollen mitosis varied among flowers and also among inflorescences. All plants were equally affected. The absence of fertile sperm cells in a considerable amount of pollen grains in this accession of B. jubata may compromise its use in breeding and could explain, at least in part, why seed production is low when compared with the amount of flowers per raceme.