Aerobic Respiration Yields about 60 Molecules of ATP per Molecule of Sucrose

The complete oxidation of a sucrose molecule leads to the net formation of four
molecules of ATP by substrate-level phosphorylation (two during glycolysis and two in the TCA
cycle),two molecules of NADH in the cytosol, and eight molecules of NADH plus two molecules of
FADH2 (via succinate dehydrogenase) in the mitochondrial matrix. Basis on measured ADP:O
values (Table 11.1), a total of approximately 28 molecules of ATP will be generated per glucose by
oxidative phosphorylation. This result in a total of 32 ATPs per glucose. (Note the difference
between this number and the value of 36 commonly seen in textbooks; the lamer value is based on
rounded-off integral values for the number of ATPs synthesized during the oxidation of each
molecule of matrix or external NADH and succinate rather than the actual measured values given
in Table 11.1.Using 50 kJ mol1 (12 kcal mol1) for the actual free energy of formation of ATP in
vivo, we find that about1606 kJ mol1 (384 kcal mol1) of free energy is conserved per mole of
glucose oxidized during aerobic respiration. This represents about 52% of the total free energy
available; the rest is lost as heat. This is a vast market improvement over the 3-5% efficiency
associated with fermentativemetabolism.
Plants Have a Cyanide-Resistant Respiration Pathway Not Found in Animals
If cyanide (I mM) is added to actively respiring animal tissues, cytochrome oxidase is
inhibited and the (respirationrate quickly drops to less than 1% of its initial level. However, must
plant tissues display a level of cyanide-resisant respiration that represents 10 -25%, but in some
tissues can be up to 100% , of the uninhibited control rate. The enzyme responsible for this oxygen
uptake has been identifed as a cyanidc-tcsistant oxidase of the plant mitochondrial electron
transport chain (Siedow and Berthold, 1986). Although the detailed mechanism of the unusual
oxidase associated with this alternave pathway has yet to be characterized, current evidence
indicates that electrons feed off the main clcctron ransport chain into the alternative pathway at the
level of the ubiquinone pool. The cyanide-resistant terminal oxidase associated with the alternative
pathway catalyzes a four electron reduction of oxygen to water and is specifically inhibited by
several compound, most notably salicylhydroxamic acid {SHAM) and n-proply gallate. Because
electrons branch to the alternative pathway from the ubquinone pool, two sites of energy
conservation (at complexes III and IV) are bypassed, and there is no evidence for an energy
conservation site on the alternative pathway between ubquinone and oxygen. Therefore, the free
energy that would normally be stored as A I P is los( as heat when electrons are shunted through
the alternative pathway.
How can such an energeticall wasteful Process as the cyanide-resistent Pathway
contributc to plant mctabolism? One example of its utility appears during floral development in

certain membranes of the Araceae (skunk cabbage Symplocarpus foetidus . Just prior to
pollination, tissue within a clublike organ on the developing floral apex (the spadix) undergoes high
rates of respiration through the alternarive Pathway, causing the spadix to heat up by as much as
14oC above ambient temperature and to volatilize odiferous compounds that attract pollinating
insects. Salieylic acid, a compound related to aspirin, has been identified as the chemical signal
responsible for initiating this the magenic event (Raskin ct al., 1989). (See box on salicylic acid in
Chapter 13).
In most plants, however, both the respiratory rates and the level of cyanide-resistant
respiration are too low to generate significant levels of heat. lt has been suggasted that the
alternative pathway might serve as an energy owerflow, oxidizing respiratory substrates that
accumulate in excess of those needed for growth, storage, or synthesis of ATP (Lambers. 1985). ln
this view, electrons "spill" into the alternative pathway only after the capacity of the main pathway is
saturated. This occurs experimentally when cyanide is added, but it could take place in vivo ii a
state 4 situation developed and the respiration rate exceeded the cell's demands for ATPT here is
evidence from research that the alternative pathway serves an energy overflow function in higher
plants (L:mbcrs, 1985)
Respiration Is Rcgulated by Energy Demandand the Concentration of Key Metabolites
The control of carbohydrate metabolism in plants is poorly understood, but it is clear that it
operates at numerous level, starting with the rate are which reduced carbon is synthesized and/or
imported into a plant tissue. In addition, because one of the major functions of respirationis to
generate ATP, the rate of ATP utilization by a cell or tissue will be of primary importance in
contfluence the level of ADP present, and this will markedly influence the rates of glycolysis-in the
cytosol and oxidative phosphorylation in thc mitochondria through mechanisms described below.
Control points appear to exist at all three stages of respiration, and only a brief overview
of some obvious features will be given here (see Douce, 1985, for-a more detailed discussion). ln
vivo, glycolysis appart to ix regulated at the level of the enzymes phosphofructokinase and
pyruvate kinase just as it is in animal respiration. Pyruvate kinase is markedly inhibited by its
product, ATP. As a result, high cytosol levels of the substrate ADP promote the pyruvate kinase
reaction, whereas high levels of ATP in the cytosol inhibit this reaction. Further, FEP, which bilds is
up in the cyrosol in response to ATP inhibitot of pyruvate kinase, is a potent inhibitor of the ATP
dependent phosphofructokine acrivity. This inhibitory effect of AT on phosphofrucrokinase is
enhanced in the presence of ATP but strongly attenuated by inorganic phosphate. These regulatory
controls allow the flow of carbon through glycolysis to respond, to a first approximation, to the cell's
energy demands by monitoring changes in the cytoplasmic IATP]/[ADP] ratio.

However, the regulation of glycolysis is more complex than outlined above. For example,
the ability of plant mitochondria to metabolize malate in addition to pyruvate (Fig. 11.4) makes ir
possible for plant cells to bypass the requlatory features of the pyruvate kinase reaction. With
phosphofructokinase, the situation is made more complex by the fact that plants can contain
appreciable levels of the ppi-dependent phosphofructokinase activity as well as the metabolic
effector fructose

2,6-bisphosphatase (fructose 2,6-P2) plays an important regulatory rolc in

glycolysis in animals, stimulating phosphofructokinase activity and inhibiting that of fructose 1,6bisphosphatase. Fructose 2,6-p2 is found I the cytosol of plant tissue, as are the enzyme fructose 6phosphate 2-kinase and fructose 2,6-bisphosphatase, which act to synthesize and break down
fructose 2,6-p2, respectively. Although the role of fructose 2,6-P2 in the regulation of plant
carbohydrate metabolism is not fully, understood, several points have been established (Huber,
l955). Fructose 2,6-P2 is present at varying levels in the cytoplasm. It markedly inhibits the activity
of fructose 1,6-bisphosphatese simulates the activity of ppi-dependent phosphofructokinase, and
little or no effect the activity of the ATP-dependent phosphofructokinase, Further information is
needed before we will fully understand the regulation at this point of the glycolytic/gluconeogenic
pathways. However, it is clear that the ppi-dependent phosphofructokinase and fructose 2.6-P2
play a role in plant carbohydrate metabolism.

The Penrose Phosphate pathway Oxidizes Glucose to Ribulose 5-phosphare and Reduces
NADH
The glycolytic pathway is nor the only route available for oxidation of glucose in the
cytosol of plant cells. The oxidativc pentose phosphate pathway (also known as the hexose
monophosphate shut) can also accomplish this (Fig. I 1.9). The first two reactions of this series
represent the oxidative events of the pathway, converting the six carbon glucose 6-phosphate to a
five carbon sugar ribulose 5-phosphate, with loss of a CO2 and generation of the molecules of
NADH (not NADH). The remaining reactions of the pathway bring about the conversion of ribulose
5-phosphatc to the glycolitic intermediates giyceraldehyde 3-phosphate and fructose 6-phosplratc,
Studies of the release of 14CO2 from isotopicallv labeled glucose indicate that glycolysis is the more
dominant pathway, accounting for 80 -95 % of the total carbon flux in most plant tissues. However
the pentose phosphate pathway does contribute, and development studies indicate that its
contribution increases as plant cells go from a meristematic to a more differentiated state (ap Rees,
1980).

The pentose phosphate pathway plays several roles in plant metabolism. The products of
the two oxidative steps are NADPH, and it is thought that this NADPH is used to drive reductive
steps associated with various biosynthetic reactions that occur in that cytosol. Such a role has
commonly been accepted for the operation of this pathway in animal tissue. However, because the
NADH dehydrogenase facing the cytosol on the mitochondrial inner membrane is also capable of
oxidizing NADPH, it is possible. that in plan 'cells some of the reducing power generated by the
pentose phosphate pathway contributes to cellular energy metabolism-that is, electrons from
NADPH may end up reducing O, and generating ATP. The pathway also produces ribose 5phosphate, a precursor of the ribose and deoxyribose needed in the synthesis of RNA and DNA.
Another intermediare on this path_way, the four-carbon erythrose 4-phosnhate combines with
PEP, in the initial reaction in the production of plant phenolic compounds, including the aromatic
amino acid and the precursors of lignin, flavonoids, and phytoalexins (chapter 13).
The pentose phosphate pathway is controlled by the initial reaction of the pathway
catalyzed by glucose-6-phosphate dehydrogenase, whose acrivity is markedly inhibited as the
NADPH:NADP ratio increases. However, measurements of the enzyme activities of the oxidative
penteso phosphate pathway in green tissues are complicated by the fact than many of the some
activities are also associated with chloroplast enzyme that catalyze the reactions of reductive
pentose phosphate pathway or Calvin cycle {Chapter 9). Both of the dehydrogenase activities of
the pentose phosphate pathway appear in chloroplasts and other plastids. This has led to
speculation that the oxidative pentose phosphate pathway might function chloroplasts under some
conditions, notably in the dart. Little operation of the oxidative pathway is likely to occur in the
chloroplast in the light, because mass action will drive the nonoxidative inteconversions of the
pathway in the direction of pentose synthesis. Moreover, glucose-6-phosphate dehydrogenase will
be inhibited in the light by the high NADPH: NADP ratio in the chloroplast as well as by a reductive
inactivation involving the ferredoxin : thioredoxin system (Chapter 9).
Respiration Is Tightly Coupled to Other Metabolic Pathways in the Cell
Although much of this chapter has focused on the role of respiration in energy
metabolism, it should be pointed out that glycolysis and the TCA cycle arc both linked to a number
important metabolic pathways, which will be covered in greater detail in Chapter 13. Glycolysis and
the TCA cycle are central to the production of a wide variety of plant metabolites, including amino
acids, lipids and related compounds, isoprenoids, and porphyrins (Fig. 1 1.10).
Not shown in Figure 11-10 are two additional reactions associated with plant
mitochondria. First' two enzyme activities found in the matrix of mitochondria from chlophylcontaining tissues, glycine decarboxylase and serine hydroxymethyltransferase, catalyze the CO2-

releasing step in photorespiration (Chapter 9). Second, plant mitochondria provide some of the
reactions associated with the breakdown of stored fats in developing seeds, which is considered
more extensively later in this chapter.
Whole-Plant Respiration
Many factors can affect the respiration rates of plant as well as the individual organs of
the plant. These include the species and growth habit of the plant, the type and age of the specific
organ, and environmettal variables such as the external oxygen concentration, temperature, and
plant water status Chapter 14). Whole-plant respiration rates, particularly when considered on a
fresh-weight basis, are generally lower than respiration rates reported for animal tissues. This is
due in large part to the considerable fraction of a mature plant cell that is metabolically inactive in
respiration (i.e., the central vacuole and the cell wall). Nonetheless, respiration rates in some plant
tissues equal the rates observed in actively respiring animal tissues (Seymour ct 1L, 1983)' so the
:plant respiratory process is not inherently slower than respiration in animals.
Even though plants generally have low respiration rates, the contribution of respiration to
the overall carbon economy of the plant can be substantial. A survey of several herbaceous
species indicated that 30 to 60% of the daily gain in photosynthetic carbon was lost to dark
respiration, although these values tended to decrease whit age of plants (Lambers 1985). Young
trees lose roughly a third of their daily photosynthate as respiration, and this loss can double in
older trees as the ratio of photosynthetic to nonphotosynthetic tissue decreases. In tropical areas,
70 to 80% of the daily photosynthetic gain can be lost to respiration owing to the high dark
respiration rates associated with elevated night temperatures.
Different Tissues and Organs Respire at Different Rates
A useful rule of thumb is that the greater the overall metabolic activity of a given tissue,
the higher its respiration rate. Developing buds usually show very high rates of respiration (on a
dry-weight basis), and respiration rates of vegetative tissues usually decrease from the growing to
more differentiated regions. In mature vegetative tissues, stems generally have the lowest
respiration rates, and leaf and root respiration varies with the plant species and the conditions
under which the plants are growing.
When a plant tissue has reached maturity, its respiration rate will either remain roughly
constant or decrease slowly as the tissue ages and ultimately senesces. An exception to this
behavior is the marked rise in respiration known as the climacteric that accompanies the onset of
ripening in many fruits (avocado, apple, banana) and senescence in detached leaves and flowers.
Both ripening and the climacteric respiratory rise are triggered by the endogenous production of

ethylene and both process can be stimulated to occur prematurely by exogenous application of
ethylene (Chapter 19). Although ripening in these fruits is associated with increased rates of protein
synthesis, which require cellular energy, the exact role of the respiratory rise is not clear. Some
fruits (citrus, pineapple, grapes) do not show a respiratory climacteric, and these fruits arc also not
generally stimulated to ripen by addition of ethylene. Many studies have shown that fruits that
undergo a climacteric respiratory rise display a high level of the cyanide-resistant pathway prior to
ripening, but apparently not during the respiratory burst itself lLzcks,1982).
Environmental Factors Can Alter Respiration Rates
The environmental factors that can affect plant respiration clearly include oxygen because
of its role as a substrate in the overall process. At 25C, the equilibrium concentration of O 2 in an
air-saturated (20% O2) aqueous solution is about 250 M. The Km value for oxygen in the
cytochrome oxidase reaction is difficult to measure exactly but is well below 1 M, so there should
be no apparent dependence of the respiration rate on external O2 concentrations (see Chapter 2
for discussion of Km). In fact, little effect on measured respiration rates is observed until the
atmospheric oxygen concentration drops below 5% for whole tissues or below 2 to 3% for tissue
slices concentration at all is due to limitations on the diffusion of oxygen through the aqueous
phase between the atmosphere

and the mitochondria. The diffusion coefficient of oxygen

dissolved in an aqueous medium is 104 less than that of oxygen diffusing through air. The fact that
atmospheric oxygen tensions as high as 5% can reduce the observed respiration rate in whole
tissues indicates that the diffusive movement of oxygen represents a true limitation on plant
respiration. Further, the existence of a diffusion-limited, aqueous pathway of oxygen movement
implies that intercellular air spaces are important in facilitating oxygen movement within plant
tissues. If there were no gaseous diffusion pathway throughout the plant, the cellular respiration
rates of many plants would be limited by an insufficient supply of oxygen.
This diffusion limitation is even more significant when the medium in which the tissue is
growing is a liquid. when plants are grown hydroponicallv it is necessary to aerate the solutions
vigorously' to keep oxygen levels high in the vicinity of the roots, and the problem of oxygen supply
also arises with plants growing in predominantly wet or flooded soils (see Chapter 14). Some
plants, particularly trees, are restricted in geographic distribution by the need to maintain a supply
of oxygen to their roots. For instance,

dogwood and tulip tree poplar can survive only in well-

drained aerated soil because their roots are unable to tolerate more than a limited exposure to a
flooded condition. Many other plants, however, can adapt to growth in flooded soils. Herbaceous
species such as rice and sunflower often rely on a network of intercellular air spaces running from
the leaves to the roots to provide a continuous, gaseous pathway for the movement of oxygen to

the flooded roots. The problem can be even moee ecute for trees that have very deep roots
growing in wet soils. Such roots mustsuryive on anaerobic (fermentative) metabolism and/or
develop structures that facilitate the movement of oxygen to the roots. Good examples to the latter
are outgrowths of the roots, called preumatophores, which protrude out of the water and provide a
gaseous pathway oxygen diffusion into the roots. Pneumatophore are found in Avicennia and
Rbizophora trees that grow in mangrove swamps under continuously floor-led conditions. Whilen
superficially similar in appearance to mangrove pneumatophores, the "knees" of cypress
(Taxodium) do not perform such a function.
Although lowering the external oxygen concentration not necessarily decrease the rate of
aerobic respiration, it does necessarily decrease the rate of carbohydrate utilization. In 1861' Louis
Pasteur discovered that yeast less glucose in the presence of air than under anaerobic conditions.
This response, known as the Pasteur effect, occurs in higher plant tissues and manifests itself as
an increased of fermentative metabolism under anaerobic conditions. In aerobic respiration,
cytosolic of ATP arc high, producing an inhibitory effect on the regulatory enzymes of the glycolytic
pathway, ATP-dependent phosphofructokinase and pyruvate kinase. In tissue maintained under
anaerobic conditions, cytosolic ATP level and its attendant inhibitory effects are decreased,
leading to higher rates of carbohydrate metabolism.
The effect of atmospheric oxygen (and temperature) on respiration is utilized in fruit
storage, where it is common to maintain fruits in the cold under conditions of 2-3yo oxygen and 35o/o CO2. The reduced temperature serves to lower the respiration rate (see below)' as does the
reduced oxygen. Low levels of oxygen arc used instead of anoxic conditions to avoid lowering
tissue oxygen tensions enough to stimulate fermentative metabolism. Carbon dioxide has little
direct effect on the respiration rate even at 3-5%, well in excess of the 0,035% normal found in the
atmosphere. High concentrations of CO2 are used in fruit storage to block the effect of the hormone
ethylene on fruit ripening (Chapter l9).
In the physiological temperature range, respiration is temperature-dependent. The
increase in respiration rate for every 10oC increase in temperature, commonly referred to as the Q10
is slightly greater than two, although in some plants an unexplained decrease in the Q10 has been
observed at temperatures above 30oC. High night temperatures are thought to account for the large
respiratory component in tropical plants, and lowered temperatures are utilized to retard
postharvest respiration rates during the storage of fruits and vegetables. However, complications
may arise from this practice. For instance, when potato tubers are stored at temperatures above
10"C, respiration and ancillary metabolic activities are sufficient to allow sprouting to produced.
Below 5oC, respiration rates and sprouting are reduced, in most tissues, but the breakdown of

stored starch and ion conversion to sucrose impart an unwanted Sweetness to the tubers. As a
compromise,

Potatoes are stored at7-9oC, which prevents the breakdown of starch while

minimizing respiration 1nd germination.

Other factors that can affect plant respiration include inorganic ions and injury. Addition
of ions to plants previously grown in distilled water stimulates respiration a phenomenon called
salt respiration. The simplest explanation for this effect is that respiratory linked ATP is required to
support the enhanced ion uptake, but other factors also seem to be involved. Physical injury to
higher plant tissues of then stimulates oxygen uptake because of increases in respiration
(mitochodria linked oxygen uptake) and in nonmitochondrial activities (e.g., lipoxygenase,
polyphenol, oxidase, peroxidase).
LIPID METABOLISM
Starch and sucrose are not the only substrates available For energy production in the
plant. Fats and oils are important storage forms of reduced carbon in many seeds, including those
of agriculturally important species such as soybean, sunflower, peanut, and cotton. Oils often serve
a major storage function in nondomesticated plants that produce small seeds. Some fruits, such as
olives and avocados, also store fats and oils.
Fats and oils belong to the general class of lipids a structurally diverse group of
hydrophobic compounds that are soluble organic solvents and highly insoluble in water. They
represent a more reduced form of carbon carbohydrates, so the that, complete oxidation of one
gram of fat or oil (-4o kJ or 9'3 kcal) cm produce considerably more ATP than the oxidation of one
gram of starch (

15.9 kJ or 379 kcal). Conversely, the biosynthesis of fats, oils, and related

molecules such as the phospholipids of membrane (see Chapter 1) requires a correspondingly

large investment of metabolic energy other lipids arc important for plant structure .and function but
are not used for energy storage. These include waxes, which make up the protective cuticle that
reduces water loss from exposed plant tissues, and terpenoids (also known as isoprenoids), a
family of compound derived from the condensation of successive five-carbon isoprene units.
lmportant terpenoids include the carotenoids involved in phorosynthesis and the sterols, which are
present in many plant membranes. The metabolism of terpenoids will be covered in greater detail in
Chapter l3.
Fats and Oils Are Triglycerides and Are Stored in Spherosomes
Fats and oils exist mainly in the form of triacylglycerols; or triglycerides, in which fatty acid
molecules are linked by ester bonds to the three hydroxyl groups of glycerol (see Figure 11.14).The
fatty acids in plants are usually straight-chain carboxylic acids having an even number of carbon
atoms. The carbon chains lengths can be as short as 12 units and as long as 20, but more

commonly 16 or 18 carbons long. Oils are liquid at room temperature, primarily due to the presence
of a number of unsaturated bonds in their component fatty acids, whereas fast are solid, having a
higher ; proportion of saturated fatty acids. The major fatty acid appearing in plant lipids are
shown in Table 11.2. The present composition of fatty acids in plant lipids varies with the species.
For example, peanut oil is about 9% palmitate acid, 59% oleic acid, and 21% linoleic acid, and
cottonseed oil is 20% palmitate acid, 30% oleic acid, and 45% linoleic acid.
Triacylglycerols in most seeds are stored in the cytoplasm of either cotyledon or endosperm cells in
organelles known as spherosomes (also called lipid bodies and oleosomes). Spherosomes have an
unusual membrane barrier that separates the triglycerides from the aqueous cytoplasm. A single
layer of phospholipids (i.e., a half bilayer) surrounds the spherosome with the hydrophilic ends of
the phospholipids exposed to the xytosol and the hydrophobic acyl side chains facing the
triacylglycerol interior. Several proteins are also located within the phospholipid half bilayer. This
unique structure appears to result from the pattern of triglyceride biosynthesis. Triglyceride
synthesis is completed by enzymes in the indoplasmic reticulum (ER), and the resulting fats
accumulate between the two monolayers of the ER membrane. bilayer. The bilayer swells apart as
more fats are put into the growing structure, and ultimately a mature spherosome buds off from the
ER (\Wanner et al., 1981).
Triacylglycerol Biosynthesis Is Energetically Expensive and Takes Place in Several Cell
Organelles
However, the acetyl CoA is first carboxylated to malonly CoA, which serves as the twocarbon donor (Fig.11.12). The reaction serves involves several enzymes, including acetyl CoA
corboxylase and a fatty acyl synthetase complex. Troughout the biosynthetic process, the growing
fatty acid remains covalently attached to a low-molecular-weight protein known as an acyl carrier
protein (ACP). The biosynthesis of fatty acids is not cheap energetically; one molecule of ATP and
two molecules of NADHP are required for the addition of each two-carbon unit.
In animals, fatty acid biosynthesis takes place in the cytosol. But in plants it is localized in
chloroplasts in leaves and in proplastids in nongreed tissue. The initial product of this series of
reactions is the C16 saturated fatty acid palmitate (16;0,.Table 11.2). A two-carbon elongation to
stearate (18;0) also takes place in the plastid (by a mechanism that does not involve the fatty acid
synthetase ..
Phospholipid Biosynthesis Occurs in the ER and mitochondria membrane
In mitochondria and most other plant membrane, phospholipids predominate, whereas in
chloroplasts the thylakoid membranes are largely composed of galactolipids (Chapter 8). The
biosynthesis of membrane lipids is complex involves several different cell compartments, and is not

completely understood in most cases (Moore, 1982). Here we will give a brief overview of the facts
that seem most well established.
Two route of phospholipids biosynthesis are recorganized within the ER, both of which
share a common pathway initially. Thase are presented in figure 1.13. At one time, it was thought
that the addition of head groups in phospholipids biosynthesis took place exclusively in the ER.
Now there is evidence that some phospholipids biosynthesis can also occur in other cell
compartments. Most notably, mitochondria contain the enzyme necessary for converting
diacylglycerol to phostphatidylcholine and phostphatidylethanolamine, and plant mitochondria are
capable of synthesizing phosphatide acid, CDP-diacylgycerol, and phostphatidylglycerol. Further,
mitochondria lener membrane lipid, cardiolipid (diphosphatidylglycerol). The extent to which
mitochondrial membranes inport lipids from the ER during organelle biogenesis is not known, nor is
the mechanism of exchange between the ER and the outer and inner mitochondrial membranes.
Phospholipids in the golgi complex, plasma membrane, and nuclear envelope (all
components of the endomembrane system that includes the ER) are probably derived from
phospholipids initially synthesized in the ER and transferred to the appropriate membrane by
vesiculation os specific regions of thr ER (Sabaniti at al.,1982). This ER- vesiculation model also
explains most of the observed features of microbody (peroxisome, glyoxysome) membrane
biogenesis (Treleease. 1984).
The syntheasis of diaciglycerol and galactolipids for the chloroplast thylakoid membranes
has been localized in spinach to a group of enzyme within the inner membrane of the chloroplast
envelope (fig.11.12) (Heemskerk and Wintermens, 1987). Synthesis of galactolipids is carried out
by an enzyme that transfer the galactosy moite from UDP-galactose diacylglycerol o from
monogalactosyldiacylglycerol. Formation of digalctosyldilglycerol involves reaction with a second
molecule of UDP-galactose.
In Germination Seeds, Lipids Are Converted into Carbohydrate
Metabolism of stored fast by oil-contained seeds involves

the conversions the

conversions of lipids to carbohydrates following germination by a process involving several different

cellular sites (Huang et al., 1983). Plants are not able to transport fats from the endosperm to the
root and shoot tissue of the germination seedling, so they must convert stored lipids to a more
mobile from of carbon, generally sucrose.
The process of converting lipid to sugar in oil seeds is triggered by germination and begins
with the hydrolysis of triglycerides stored in the spherosomes to free fatty acids, followed by
oxidative breakdown of the fatty acids to produce acetyl CoA (Fig .11.14). The Katter reaction
takes place in a type of microbody called glycoxysomes, organelles bounded by a single

membrane that are found in the oil-rich storage tissue of seed. The discovery of glycoxysomes in
castor beans and much of ours understading of glyoxysomel fuction are due to the work of harry
Beevers and his colleagues at Purdue University and the university of California, Santa Cruz (see
Beevers, 1990). Acetyl CoA is metabolized in the glycoxysome via the glyxylate cycle (fig. 11.14) to
produce succinate, which is transported from the glycoxysome to the mitochondria, where it is
further metabolized in the glycoxysome. The process ends in the cytosol with the conversion of
oxaloacetate to glucose via gluconeogenesis. Although some of this carbon is diverted to others to
other metabolic reactions in certain oil seeds, in castor baen the process is so efficient that each
gram of lipid metabolized results in the formation of one gram of carbohydrate, equivalent to 40%
recovery of free energy in the form of carbon bonds.
The initial step in the conversions of lipids to carbohydrate involves the breakdown of
triglycerides stored in the spherosomes by the enzyme lipase, which, at least in castor bean, is
located on the half membrane that serves as the spherosome outer boundary. The lipase
hydrolyzes triglycerides to three molecules of fatty acid and glycerol. Corn and cotton also contain
a lipase activity in the spherosome, but peanut, soybean, and cucumber show lipase activity in the
glycoxysome instead. During the breakdown of lipids, there is generally a close physical
association between spherosomes and glycoxysomes.
Following hydrolysis, triglycerides, the resulting fatty acids enter the glyoxysome , where
they are activated to fatty acyl CoA by the enzyme fatty-acyl-CoA sythase. Fatty acyl CoA is the
initial substrate for the -oxidation series of reactions in which Cn fatty acids are sequentially broken
down to n/2 molecules of acetyl CoA. This reaction sequence is similar to that associated with the
breakdown of fatty acids in animal tissue an involves the reducation of 1/2 O 2 to H2O and the
formation of one NADH for each acetyl CoA produced. In mammalian tissue, the four enzyme
activites associated with -oxidation appear in the mitochondria, whereas in plant seed storage
tissue they are exclusively localized in the glyoxysome. Interestingly, in plant vegetative tissue
(e.g., mung been hypocotil, potato tuber), the -oxidation reactions are localized in a related
organelle, the peroxisome, rather than in the mitochondria (Gerhardt, 1983), suggesting that the
limited breakdown of lipids that does occur in plant nonstorage tissues takes place in peroxisomes
and not mitochondria.
The acetyl CoA produced -oxidation is further metabolized in the glyoxysome through a
series of reactions that make up the glyoxylate cycle (Fig.11.4). Initially, the acetyl CoA reach with
oxaloacetate to give citrate, which it turn undergoes isomerization to isocitrate. Both of the enzyme
involved here (citrate synthase and aconitase) catalyze the some reaction as in the TCA cycle in
the mitochondria. The next two reactions are unique to the glyoxylate pathway. First, isocitrate (C6)

is cleaved by the enzyme isocitrate lyase to give succinate (C4) and glyoxylate (C2). Next, malate
synthase combines a second molecule of acetyl CoA with glyoxylate to produce malate. According
to the classical view of the operation of the glyoxlate, the malate is then oxuidized by malate
dehydrogenase to oxaloacetate, which can combine with another acetyl CoA to continue the cycle.
The function on the glyoxylate cycle is, in essence, to convert two moleculesof acetyl CoA
to succinate. The succinate that moves to mitochondria, where it is converted to malate by the
normal TCA cycle reactions. The resulting malate can be moved out of the mitochondria in
exchange for the incoming succinate via the dicarboxylate transporter. The malates is then oxidized
to oxaloacetate by cytosolic malate dehydrogenase, and the resulting oxaloacetate is converted to
carbohydrate. This conversions requires circumventing the irreversibility of the pyruvate kinase
reaction and is facilitated by the enzyme PEP carboxynase, which utilizes the phosphorylating
ability of ATP to convert oxaloacetate to PEP and CO2. From PEP, gluconeogenesis can procced
to the production of glucose, as outline on page 268. Sucrose in the ultimate product of this
process, being the primary from in which reduced carbon is translocated from the cotyledons to the
growing seedling tissue.
Although the pathway for the mobilization of triglycerides has been best characterized in
castor been (ricinus communities), it seems to be similar in the storage tissue of other oil seeds.
However, not all seeds carry out a quantitative conversions of fat to sugar. In castors bean, the
endosperm degenarates after the fat and protein reserves are fully utilized. In many oil seeds, such
as sunflower (helianthus annus), cotton (gossipium birsutum), differentiate into actively
photosynthesizing organs after the food reserves are used up. In these tissues only part of the
stored lipid is converted to exported carbohydrate. Much of the lipid-derived carbon remains in the
cotyledos, where it contributed to the synthesis of chloroplast and other cellular structures. As the
greening process takes place, there is a transition in the microbody population of these cells that
shows fewer of the characteristics of glyoxysomes and more of those of leaf type peroxisomes.
Such a transition is in keeping with the decreased requirement for the breakdown of stored lipids
and the increased need to metabolize the pruducts of photorespiration as the tissue goes from a
heterotrophic to a more autotrophic mode of metabolism.
Summary
Higher plant respiration couples the complete oxidation of reduced cellular carbon
generated during photosynthesis to the synthesis of ATP. Respiration takes place in three stages
glycolysis, the tricarboxylic acid cycle, and the electron transport chain. In glycolysis, carbohydrate
is converted in the cytosol to pyruvat with the synthesis of a small amount of ATP via substratelevel phosphorylation. Pyruvat is subsequently oxidized within the mitochondrial matrix through the

TCA cycle, generating a large number of reducing equivalents in the form of NADH and FADH2. In
the third stage, NADH and FADH2 are oxidized by the electron transport chain, which is associated
with the inner mitochondrial membrane. The free energy released during electron transport is used
to synthesize a large amount of ATP in a process known as oxidative phosphorylation. ATP
synthesis is accomplished by the generation of a proton gradient across the inner mitochondrial
membrane during electron transfer. The FO-F1-ATP synthase complex then couples the break-down
of the proton gradient to the conversion of ADP and Pi to ATP.
The regulation of substrate oxidation during respiration involves control points at
glycolysis, the TCA cycle and the electron transports chain. Aerobic respiration in higher plants has
several unique features, including the presence

of cyanide-resistent oxidative pathway.

Carbohydrates can also be oxidized via the pentose phosphate pathway, in which the reducing
power generated produces NADPH for byosinthetic purposes. Numerous glycolytic and TCA cycle
reactions also provide the starting points for different byosinthetic pathways.
Many factors can affect the respiration rate observed at the whole-plant level. These
include and age of the plant tissue and such environmental factors as the external oxygen
concentration and temperature. Because respiration rates contribute to the overall net carbon
balance of a plant, variations in whole-plant respiration rates can affect agronomic yields.
Lipids play a major role in higher plants; amphipathic lipids serves as the primary non
protein components of the membranes, while triglycerides (fats and oils) are an efficient storage
from a reduced carbon, particularly in seed. Triglycerides are stored in subcellular organelles
known as spherosomes. Synthesis of the long-chain fatty acids that makes up triglycerides takes
place in the plastids of higher plants, and the conversions of the fatty triglycerides occurs on the
membranes of the endoplasmic reticulum. During generation in seed the store triglycerides, the
stored lipids are metabolized to carbohydrates in a series of reactions that involve a metabolic
sequence known as the glyoxylate cycle. This cycle takes place in organelles called glyoxysomes.
The reduced carbon generated during lipid breakdown in the glyoxysomes is ultimately converted
to carbohydrate in the glyoxysomes is ultimately converted to carbohydrate in the cytosol by the
process known as gluconeogenesis.