Reproductive behavior of Acroceras macrum Stapf

1,

Ferrari Usandizaga S.C. Schedler M. , Ledesma D. , Martnez E.J.

2,3

, Acua C.A.

2,3

1: Agricultural and livestock experimental station Corrientes, INTA. 2:IBONE-CONICET. 3: School of Agrarian Sciences, UNNE. efeseis@yahoo.com.ar

Introduction
Acroceras macrum is a warm-season grass used as forage and it is
characterized for its high nutritive value. The low seed yield obligates the
farmers to spread it in a vegetative way, limiting its adoption as forage.
There is a serious need of information about the reproductive behavior of
this species to initiate a breeding program. A plant collection accomplished
in northeastern Argentina resulted in 22 tetraploid (2n=4x=36) and 5
hexaploid (2n=6x=54) genotypes. The objective of this research was to
determine the mode of reproduction in that collection.

Materials and Methods

Clarified ovaries with methylsalicylate at different stages of development
were observed using differential interference contrast microscopy to study
the characteristics of the embryo sacs. Seed yield was studied in open and
self-pollination. Cross-pollination between different genotypes with
variables ranges of genetic distances also was performed (Fig. 1) and the
hybrid origin of the seed obtained was tested with ISSR molecular
markers.

Figure 1. Acroceras macrum experimental plot for the evaluation of seed

production in terms of self-pollination, open pollination and controlled crosses
between different genotypes.

Results
Megasporogenesis showed a developmental state according with the
normal meiotical pattern in both cytotypes (Fig. 2 a-h). All observed
embryo sacs had the Polygonum structure typical of Paniceae, with an egg
cell, two synergids, a binucleated central cell and 3 to 7 antipodals (Fig. 2
i-k). The embryo sacs types observed in mature ovules are resumed in
Table 1. Seed set in open-pollinated from tetraploid genotypes varied from
3.7 to 69%, with an average of 34.5% (Fig. 3). From open-pollinated
hexaploid genotypes it varied from 0.2 to 3.7%, and 1.5% in average (Fig.
3). Seed set in self-pollination from tetraploid genotypes varied from 0.3 to
18%, with an average of 5.1%. Hexaploid genotypes did not produced
seeds in self-pollination (Fig. 3). Cross combination produced variable seed
set depending on the genotype combination and the cytotype of each
parent. When the mother was an hexaploid, crosses did not result in fertile
seeds (Table 2). Crosses between tetraploids genotypes produced several
plants. From these crosses, between 87.5% to 100% were hybrids.

Figure 2. Megasporogenesis and megagametogesis in A. macrum. a. Megaspore

Mother Cells (MMCs) (arrow). b. Megaspores produced at meiosis from the MMC
(arrow). c. Deteriorated megaspores at micropillar end (arrow). d. Functional
megaspore (arrow) and remains of deteriorated megaspores. e. Binucleate stage
(functional megaspore 1st mitosis ). f. Tetranucleate stage (2nd mitosis) and
polarization of nuclei (arrows). g. Nuclei grouped at micropylar pole (arrow). h, i,
j, k. Meiotic embryo sacs (arrow: binucleate central cell). h. Mature embryo sac.
i. Polygonum sac. j. Nuclear proliferation of antipodes. k. Meiotic embryo sac
with five antipodes. * an: antipodes; o: egg cell.
Table 1. Embryo sacs types observed in mature ovules from tetraploid (4x) and
hexaploid (6x) cytotypes of A. macrum.

Cytotype

Aborted sacs
(%)

No visible
ontogenetic
structures on sac
(%)

Meiotically
derived mature
sacs
(%)

Immature
structures at
anthesis
(%)

4x

8a*

9a*

23a*

1a*

6x

22b*

11a*

28a*

19b*

*: no significant differences between the studied genotypes were observed within each cytotype

4x open pollination
4x self pollination

6x open pollination
6x self pollination

Figure 3. Seed set from 27 genotypes of A. macrum.

Table 2. Seed set from different crossing types between tetraploid (4x) and
hexaploid (6x) genotypes of A. macrum.

Conclusions
Our results are according to sexual reproductive behavior and allogamy
due to self-incompatibility for both tetraploid and hexaploid cytotypes.

Crossing type

x
4x-4x
4x-6x
6x-4x
6x-6x

Average seed set

(%)

Rank
(%)

15.1a
3.9b
0b
0b

0-39.5
1.2-10.5
0
0