Leaf Abscission in Soybean: Cytochemical and Ultrastructural Changes following Benzylaminopurine Treatment

6-benzylaminopurine (BAP) delays leaf abscission of soybeanGlycine max (L.) Merr. Abscission of the distal pulvinus ofprimary leaves was induced in 12-d-old seedlings or explantsby removal of the leaf blade. BAP applied to the cut end ofthe pulvinus following leaf blade removal delayed abscission.Discoloration of the pulvinus occurred before abscission commencedand the number of grana in chloroplasts within cortical parenchymacells of the pulvinus decreased over time following leaf bladeremoval. BAP prevented discoloration of pulvinus tissues anda decrease in grana number. Starch grains within amyloplastsof cells of the starch sheath in the pulvinus disappeared followingleaf blade removal, whereas starch accumulated within the abscissionzone prior to abscission. BAP prevented this apparent redistributionof starch and instead promoted an increase in starch withinplastids of cortical parenchyma cells of the pulvinus. Duringthe abscission process, cells within the separation layer enlargedand their nuclei and nucleoli became more evident prior to theirseparation from one another. Cell separation resulted from breakdownof middle lamellae and partial degradation of primary cell walls.Cycloheximide applied directly to the external surface of theabscission zone inhibited abscission in a similar way to theBAP treatment. These results suggest that BAP prevents abscissionby altering patterns of starch distribution in the pulvinusand abscission zone and by inhibiting the synthesis of proteinsthat typically appear de novo in induced abscission zone tissues. Key words: Benzylaminopurine, BAP, Soybean, Pulvinus, Abscission, amyloplast.     

Early Endosperm Development in Ovules of Soybean, Glycine max (L) Merr. (Fabaceae)

Endosperm development was studied in normally setting flowersand pods of soybean from anthesis to a pod length of 10–20mm. The free-nuclear stage following double fertilization istypified by loss of starch and increasing vacuolation. The cytoplasmprovides evidence of extensive metabolic activity. Wall ingrowths,already present at the micropylar end of the embryo sac wallprior to fertilization, develop along the lateral wall of thecentral cell as well as at the chalazal endosperm haustorium.Endosperm cellularization begins when the embryo has developeda distinct globular embryo proper and suspensor. Cellularizationstarts at the micropylar end of the embryo sac as a series ofantidinal walls projecting into the endosperm cytoplasm fromthe wall of the central cell. The free, growing ends of thesewalls are associated with vesicles, microtubules, and endoplasrnicreticulum. Pendinal walls that complete the compartmentalizalionof portions of the endosperm cytoplasm are initiated as cellplates formed during continued mitosis of the endosperm nuclei.Endosperm cell walls are traversed by plasmodesmata. This studywill provide a basis for comparison with endosperin from soybeanflowers programmed to abscise. Glycine max, soybean, endosperm, ovules     

Sieve-Element Ontogeny in the Aerial Shoot of Equisetum hyemale L.

The aerial shoots of Equisetum hyemale L. var. affine (Engelm.)A. A. Eat. were examined with the electron microscope as partof a continuing study of sieveelement development in the lowervascular plants. Young E. hyemale sieve elements are distinguishablefrom all other cell types within the vascular system by thepresence of refractive spherules, proteinaceous bodies whichdevelop within dilated portions of the endoplasmic reticulum(ER). Details of cell wall thickening differ between protophloemand metaphloem sieve elements. Following cell wall thickeningthe ER increases in quantity and aggregates into stacks. Shortlythereafter, nuclear degeneration is initiated. During the periodof nuclear degeneration some cytoplasmic components-dictyosomes,microtubules and ribosomes-degenerate and disappear, while organellessuch as mitochondria and plastids persist. The latter undergostructural modifications and become parietal in distribution.Eventually the massive quantities of ER are reduced, leavingthe lumen of the cell clear in appearance. At maturity the plasmalemma-linedsieve element contains a parietal network of tubular ER, aswell as mitochondria, plastids, and refractive sphemh At thistime many of the spherules are discharged into the region ofthe wall. Sieveelement pores occur in both lateral and end walls.At maturity many pores are traversed by large numbers of ERmembranes. The metaphloem sieve elements of the mid-internodalregions apparently are sieve-tube members. The connections betweenmature protophloem sieve elements and pericycle cells are associatedwith massive wall thickenings on the pericyclecell side.     

A Flower and Pod Staging System for Soybean

Flower and pod abscission limit soybean yield. A system forquantifying flower and pod development based on the morphologicalappearance of the flower prior to and following anthesis hasbeen developed to aid in studies of pod abscission. Changesin the appearance of the corolla, primarily the banner petal,are used to distinguish the different stages of the system.External pistil dimensions have been correlated with internalfeatures for each stage of development. From anthesis to podset, pistil length and weight increase almost two- and fivefold,respectively, and ovule development progresses from unfertilizedegg cells to embryos surrounded by cellular endosperm. Pod determinedare correlated with ovule length and width and embryo cell number.Flower and pod stages can be determined in situ, thus permittingnon-destructive observation and experimental manipulation offlowers or pods without necessarily impeding their development.Stages have been identified that indicate precisely when podset occurs and when young pods cease growing and ultimatelyabscise. This system of flower and pod staging is useful instudies designed to assess effects of abiotic or biotic stressand genetic factors on pod set and abortion. Abscission, anthesis, Glycine max (L.) Merr, embryo development, pod set     

Changes in Soybean Raceme and Petiole Anatomy Induced by 6-Benzylaminopurine

An enlargement of the peduncle and rachis of the terminal racemeand the petiole of the uppermost mainstem leaf was observedin soybean plants [Glycine max (L.) Merr.] treated with thecytokinin, 6-benzylaminopurine (BAP). Histological studies wereperformed to determine the timing and extent of anatomical changesaccompanying BAP treatment. Swelling of treated ‘Tracy-M’peduncles, rachises, and petioles was observed within 4–6d after treatment initiation. A significant increase in totalcross-sectional tissue area was observed at lower and intermediateinternodes of treated rachises after 11 d. Rachis enlargementwas due to increases in both cell size and cell number, particularlyof the vascular tissue. In treated petioles of IX93-100, procambialcells of vascular bundles were the first to respond to the BAPtreatment. These cells differentiated into a vascular cambiumwhich formed secondary xylem and phloem. Soybean, Glycine max (L.) Merr., anatomy, rachis, BAP.     

Ultrastructural Changes of the Egg Apparatus Associated with Fertilization and Proembryo Development of Soybean, Glycine max (Fabaceae)

As part of a study involving pod retention in soybean, Glycinemax (L.) Merr., we investigated changes occurring in the eggapparatus of non-abscised flowers from the time immediatelypreceding fertilization through early embryogeny. Prior to theentry of the pollen tube into the embryo sac, one of the synergidsbegins to degenerate as evidenced by increased electron densityand a loss of volume. This cell serves as the site of entryfor the pollen tube. The cytoplasm of the second, or persistentsynergid, remains unaltered until after fertilization. Bothsynergids contain, in addition to a filiform apparatus, a singleunidentified inclusion of flocculent material located in thechalazal portion of each cell. The zygote can be distinguishedfrom the egg by its consistently narrow wall; and it dividesto form a proembryo, a mass of cells not yet differentiatedinto embryo proper and suspensor. The basal cells of the proembryoare more vacuolate than the apical ones, characteristic of thebasal vacuolation of both egg and zygote. Cells of the proembryoare connected to one another via plasmodesmata, and with theexception of the basal-most cell, are isolated symplasticallyfrom the surrounding endosperm. Wall ingrowths frequently occurin certain cells of the proembryo, notably those cells in contactwith the degenerate synergid and embryo sac wall. At a laterstage of ontogeny, by which time the globular embryo properhas become distinct from the suspensor, the wall ingrowths areconcentrated in the suspensor. Glycine max, soybean, embryogeny, synergids