4

Plants and Inorganic Nutrients

Unlike heterotrophic organisms, which depend for their existence on energy-rich organic molecules previously synthesized by other organisms, plants must survive in an entirely inorganic environment. All organisms must continually draw material substance from their environment in order to maintain their metabolism, growth, and development. The means for making these materials available to the organism is the sub ect of plant nutrition. !lant nutrition is traditionally treated as two separate topics" organic nutrition and inorganic nutrition. #rganic nutrition focuses on the production of carbon compounds, specifically the incorporation of carbon, hydrogen, and oxygen via photosynthesis, while inorganic nutrition is concerned primarily with the ac$uisition of mineral elements from the soil.

4.1.2 THE USE OF HY !OPONI" "U#TU!E HE#PE TO EFINE THE $INE!%# !E&UI!E$ENTS OF P#%NTS
%. &achs h'droponic culture 'growing plants in a defined nutrient solution(, The nutrient solution devised by &achs contributed a total of nine mineral nutrients '), *, !, +a, &, *a, +l, ,e, -g(. +arbon, hydrogen, and oxygen were excluded from this total because they were provided in the form of carbon dioxide and water and were not considered mineral elements. *ote that the solution must be aerated in order to obtain optimal root growth and nutrient uptake. A solution that is not aerated becomes depleted of oxygen, a condition known as ano(ia. Anoxia inhibits the respiration of root cells and, because nutrient uptake re$uires energy, reduces nutrient uptake.

4.2 THE ESSENTI%# NUT!IENT E#E$ENTS

4.2.1 SE)ENTEEN E#E$ENTS %!E EE$E TO *E ESSENTI%# FO! P#%NT +!O,TH %N E)E#OP$ENT
-ost plants re$uire a relatively small number of nutrient elements in order to successfully complete their life cycle. Those that are re$uired are deemed to be essential nutrient ele-ents. .ssentiality is based primarily the" '/( in its absence the plant is unable to complete a normal life cycle 'if a plant is unable to produce viable seed when deprived of that element, then that element is deemed essential.( '0( that element is part of some essential plant constituent or metabolite . '1( an essential element must act directly in the metabolism of the plant

4.2.2 THE ESSENTI%# NUT!IENTS %!E +ENE!%##Y "#%SSE $%"!ONUT!IENTS O! $I"!ONUT!IENTS

%S EITHE!

The distinction between macro- and micronutrients simply reflects the relative concentrations found in tissue or re$uired in nutrient solutions and does not infer importance relative to the nutritional needs of the plant. .lements classified as macro '2( and micro'3( has to be found to be re$uired greater or less than /4 mmole kg5/ of dry weight. *ickel is an essential component of urease. Urease catalyzes the hydrolysis of urea into *6 1 and +#0 .

4.. *ENEFI"I%# E#E$ENTS

7n addition to the /8 essential elements some plants 'lower plants, not higher( appear to have additional re$uirements.

4...1 SO IU$ IS %N ESSENTI%# $I"!ONUT!IENT FO! "4 P#%NTS

A sodiu- re$uirement was first demonstrated for the bladder salt-bush ' Atriplex vesicaria(,

!lants grown in the depleted solution showed reduced growth, chlorosis 'yellowing due to loss of chlorophyll(, and necrosis 'dead tissue( of the leaves. sodium is generally essential as a micronutrient for plants utilizing specifically the +9 photosynthetic pathway, but not for most +1 plants.

4...2 SI#I"ON $%Y *E *ENEFI"I%# FO! % )%!IETY OF SPE"IES

. &ilicon is seen in maize 'Zea mays( and scouring rush 'Equisetum arvense(, &ilicon seems to be particularly beneficial to grasses, where it accumulates in the cell walls, especially of epidermal cells, and may play a role in fending off fungal infections or preventing lodging, a condition in which stems are bent over by heavy winds or rain.

4.... "O*%#T IS !E&UI!E *Y NIT!O+EN/FI0IN+ *%"TE!I%

+obalt is essential for the growth of legumes, which are hosts to symbiotic nitrogen-fixing bacteria. 7n this case, the re$uirement can be traced to the needs of the nitrogen-fixing bacterium rather than the host plant. 7n addition, when legumes are provided with fixed nitrogen such as nitrate, a cobalt re$uirement cannot be demonstrated.

4...4 SO$E P#%NTS TO#E!%TE HI+H "ON"ENT!%TIONS OF SE#ENIU$

legume genus Astragalus 'milk-vetch or poison-vetch(

4.4.1 % P#%NT1S !E&UI!E$ENT FO! % P%!TI"U#%! E#E$ENT IS EFINE IN TE!$S OF "!ITI"%# "ON"ENT!%TION
nutrient levels below the critical concentration, that nutrient becomes limiting to growth. :hen nutrient levels exceed the critical concentration, that nutrient is, with one $ualification, no longer limiting.

4.4.2 NIT!O+EN IS % "ONSTITUENT OF $%NY "!ITI"%# $%"!O$O#E"U#ES

plants absorb nitrogen from the soil solution primarily as inorganic nitrate ion '*# 1 ( and '*6;9 ( ion. incorporated into amino acids, proteins, and other nitrogenous organic molecules. *itrogen is most often limiting in agricultural situations. proteins, nucleic acids, certain hormones 'e.g., indole-1-acetic acid< cytokinin(, and chlorophyll. symptoms of nitrogen deficiency are a slow, stunted growth and a general chlorosis of the leaves. *itrogen is very mobile in the plant. As the older leaves yellow and die, the nitrogen is mobilized exported from the older leaves to the younger, more rapidly developing leaves. +onditions of nitrogen stress will also lead to an accumulation of anthocyanin pigments in many species, contributing a purplish color to the stems, petioles, and the underside of leaves.

4.4.. PHOSPHO!OUS IS P%!T OF THE NU"#EI" %"I *%"2*ONE %N H%S % "ENT!%# FUN"TION IN INTE!$E I%!Y $ET%*O#IS$
!hosphorous is available in the soil solution primarily as forms of the polyprotic phosphoric acid '6 1!#9(. phosphorous, rather than nitrogen, is most commonly the limiting element in natural ecosystems. #ne of the more successful strategies developed by plants for increasing the uptake of phosphorous is the formation of intimate associations between roots and soil fungi, called mycorrhiza.

4.4.4 POT%SSIU$ %"TI)%TES EN3Y$ES %N FUN"TIONS IN OS$O!E+U#%TION

!otassium ');( is the most abundant cellular cation and so is re$uired in large amounts by most plants. 7n agricultural practice, potassium is usually provided as potash 'potassium carbonate, )0+#1(. !otassium is fre$uently deficient in sandy soils because of its high solubility and the ease with which ); leaches out of sandy soils. !otassium is an activator for a number of enzymes, most notably those involved in photosynthesis and respiration. &tarch and protein synthesis are also affected by potassium deficiency. !otassium serves an

45 "hapter 4 6 Plants and Inorganic Nutrients

important function in regulating the osmotic potential of cells '+hapter 1(. As an osmoregulator, potassium is a principal factor in plant movements, such as the

opening and closure of stomatal guard cells '+hapter 3( and the sleep movements, or daily changes in the orientation of leaves '+hapter 01(. =ecause it is highly mobile, potassium also serves to balance the charge of both diffusible and nondiffusible anions. Unlike other macronutrients, potassium is not structurally bound in the plant, but like nitrogen and phosphorous is highly mobile. >eficiency symptoms first appear in older leaves, which characteristically develop mottling or chlorosis, followed by necrotic lesions 'spots of dead tissue( at the leaf margins. 7n monocotyledonous plants, especially maize and other cereals, the necrotic lesions begin at the older tips of the leaves and gradually progress along the margins to the younger cells near the leaf base. &tems are shortened and weakened and susceptibility to root-rotting fungi is increased. The result is that potassium-deficient plants are easily lodged.

4.4.7 SU#FU! IS %N I$PO!T%NT "ONSTITUENT OF P!OTEINS8 "OEN3Y$ES8 %N )IT%$INS

&everal forms of sulfur are found in most soils, including iron sulfides and elemental sulfur. &ulfur is taken up by plants, however, as the divalent sulfate anion '&# 05 9 (. &ulfur deficiency is not a common problem because there are numerous microorganisms capable of oxidizing sulfides or decomposing organic sulfur compounds. 7n addition, heavy consumption of fossil fuels in industry as well as natural phenomena such as geysers, hot sulfur springs, and volcanos together contribute large amounts of sulfur oxides '&#0 and &#1( to the atmosphere. 7ndeed it is often difficult to demonstrate sulfur deficiencies in greenhouses in industrial areas because of the high concentrations of airborne sulfur. &ulfur is particularly important in the structure of proteins where disulphide bonds '?&?&?( between neighboring cysteine and methionine residues contribute to the tertiary structure, or folding. &ulfur is also a constituent of the vitamins thiamine and biotin and of coenzyme A, an important component in respiration and fatty acid metabolism. 7n the form of iron-sulfur proteins, such as ferredoxin, it is important in electron transfer reactions of photosynthesis and nitrogen fixation. The sulfur-containing thioc'anates and isothioc'anates 'also known as mustard oils( are responsible for the pungent flavors of mustards, cabbages, turnips, horseradish, and other plants of the family =rassicaceae. =ecause of the presence of mustard oils, many species of the =rassicaceae prove fatal to livestock that graze on them. -ustard oils also appear to serve as a defense against insect herbivory. &ulfur deficiency, like nitrogen, results in a generalized chlorosis of the leaf, including the tissues surrounding the vascular bundles. This is due to reduced protein

synthesis rather than a direct impairment of chlorophyll synthesis. 6owever, chlorophyll is stabilized by binding to protein in the chloroplast membranes. :ith impaired protein synthesis, the ability to form stable chlorophyll-protein complexes is also impaired. Unlike nitrogen, however, sulfur is not readily mobilized in most species and the symptoms tend to occur initially in the younger leaves.

4.4.9 "%#"IU$ IS I$PO!T%NT IN "E## I)ISION8 "E## % HESION8 %N %S % SE"ON $ESSEN+E!

+alcium is taken up as the divalent cation '+a0;(. +alcium is abundant in most soils and is seldom deficient under natural conditions. +alcium is important to dividing cells for two reasons. 7t plays a role in the mitotic spindle during cell division and it forms calcium pectates in the middle lamella of the cell plate that forms between daughter cells. 7t is also re$uired for the physical integrity and normal functioning of membranes and, more recently, has been implicated as a second messenger in a variety of hormonal and environmental responses. As a second messenger involved in protein phosphorylation, +a0; is an important factor in regulating the activities of a number of enzymes. =ecause of its role in dividing cells, calcium deficiency symptoms characteristically appear in the meristematic regions where cell division is occurring and new cell walls are being laid down. @oung leaves are typically deformed and necrotic and, in extreme cases, death of the meristem ensues. 7n solution cultures, calcium deficiency results in poor root growth. The roots are discolored and may feel AAslipperyBB to the touch because of the deterioration of the middle lamella. +alcium is relatively immobile and the symptoms typically appear in the youngest tissues first.

4.4.4 $%+NESIU$ IS % "ONSTITUENT OF THE "H#O!OPHY## $O#E"U#E %N %N I$PO!T%NT !E+U#%TO! OF EN3Y$E !E%"TION
Cike calcium, magnesium is also taken up as the divalent cation '-g0;(. -agnesium is generally less abundant in soils than calcium but is re$uired by plants in relatively large amounts. -agnesium deficiencies are most likely in strongly acid, sandy soils. -agnesium has several important functions in the plant. =y far the largest proportion is found in the porphyrin moiety of the chlorophyll molecule, but it is also re$uired to stabilize ribosome structure and is involved as an activator
4.4 Nutrient Functions and Deficiency Symptoms 41

for numerous critical enzymes. 7t is critical to reactions involving AT!, where it serves to link the AT! molecule to the active site of the enzyme. -g0; is also an activator for both ribulosebisphosphate carboxylase and phosphoenolpyruvate carboxylase, two critical enzymes

in photosynthetic carbon fixation '+hapters 3 and /D(. The first and most pronounced symptom of magnesium deficiency is chlorosis due to a breakdown of chlorophyll in the lamina of the leaf that lie between the veins. +hloroplasts in the region of the veins are for some reason less susceptible to magnesium deficiency and retain their chlorophyll much longer. -agnesium is also $uite mobile. 7t is readily withdrawn from the older leaves and transported to the younger leaves that are more actively growing and synthesizing chlorophyll. +onse$uently, chlorosis due to -g0; deficiency is, at least initially, most pronounced in the older leaves.

4.4.: I!ON IS !E&UI!E FO! "H#O!OPHY## SYNTHESIS %N E#E"T!ON T!%NSFE! !E%"TIONS

#f all the micronutrients, iron is re$uired by plants in the largest amounts 'it is considered a macronutrient by some(. 7ron may be taken up as either the ferric ',e 1;( or ferrous ',e0;( ion, although the latter is more common due to its greater solubility.1 The importance of iron is related to two important functions in the plant. 7t is part of the catalytic group for many redox enzymes and it is re$uired for the synthesis of chlorophyll. 7mportant redox enzymes include the heme-containing cytochromes and non-heme iron-sulfur proteins 'e.g., Eieske proteins, ferredoxin, and photosystem 7( involved in photosynthesis '+hapter 8(, respiration '+hapter /4( and nitrogen fixation '+hapter //(. >uring the course of electron transfer the iron moiety is reversibly reduced
7n the scientific literature, particularly that body of literature dealing with iron uptake by organisms, the ferric form of iron is also referred to as ,e'777( 'iron-three(. =y the same convention, ferrous iron is referred to as ,.'77( 'iron-two(.
1

from the ferric to the ferrous state. 7ron is also a constituent of several oxidase enzymes, such as catalase and peroxidase. 7ron is not a constituent of the chlorophyll molecule itself and its precise role in chlorophyll synthesis remains somewhat of a mystery. There is, for example, no definitive evidence that any of the enzymes involved in chlorophyll synthesis are iron-dependent. 7nstead, the iron re$uirement may be related to a more general need for iron in the synthesis of the chloroplast constituents, especially the electron transport proteins. 7ron deficiencies invariably lead to a simultaneous loss of chlorophyll and degeneration of chloroplast structure. +hlorosis appears first in the interveinal regions of the youngest leaves, because the mobility of iron in the plant is very low and it is not easily withdrawn from the older leaves. +hlorosis may progress to the veins and, if the deficiency is severe enough, the very small leaves may actually turn white. 7ron deficiencies are common because of the propensity of ,e1; to form insoluble hydrous oxides

',e0#1F160#( at biological p6. This problem is particularly severe in neutral or alkaline calcareous soils. #n the other hand, iron is very soluble in strongly acidic soils and iron toxicity due to excess iron uptake can result. The problem of iron deficiency can usually be overcome by providing chelated iron, either directly to the soil or as a foliar spray. A chelate 'from the Greek, chele or claw( is a stable complex formed between a metal ion and an organic molecule, called a chelating agent or ligand. The ligand and the metal ion share electron pairs, forming a coordinate ;ond. =ecause chelating agents have a rather high affinity for most metal ions, formation of the complex reduces the possibility for formation of insoluble precipitates. At the same time, the metal can be easily withdrawn from the chelate for uptake by the plant. #ne of the more common synthetic chelating agents is the sodium salt of eth'lenedia-inetetraacetic acid <E T%= ',igure 9.9(, known commercially as versene or sequestrene. .>TA and similar commercially available chelating agents, however, are not highly specific and will bind a range of cations, including iron, copper,
FI+U!E 4.4 E(a-ples o> organic acids that >unction as chelating agents. Eth'lenedia-inetetraacetic acid <E T%= is a s'nthetic acid in co--on co--ercial use. "o-ple(ed ?ith iron8 it is sold under the trade na-e )ersenate. "a>>eic acid is one o> se@eral naturall' occurring phenolic acids that -a' ;e secreted ;' roots.

* +60 +60 * ##+ 60+ +60+## ##+ 60+ +60+##

Ethylenediamine tetraacetic acid (EDTA)

6# # 6 6+ + +## 6
Caffeic acid
H H H H H

42 "hapter 4 6 Plants and Inorganic Nutrients

zinc, manganese, and calcium. *atural chelating agents, including porphyrins 'as in hemoglobin, cytochromes, and chlorophyll( and a variety of organic and phenolic acids, are far more specific for iron. The importance of iron in plant nutrition is highlighted by the strategies plants have developed for uptake under conditions of iron stress. 7ron deficiency induces several morphological and biochemical changes in the

roots of dicots and nongraminaceous monocots. These

ATP ADP + Pi

!hiAosphere $e-;rane "'tosol A. B.

H+ Fe777 Fe777 Fe777
Soil colloid

Fe777 Fe777 Fe777 Fe77 Fe77

Fe777 Reductase

Ch Ch PS PS PS PS Fe777 Fe777

FI+U!E 4.7 T?o strategies >or the solu;iliAation and uptaBe o> sparingl' solu;le inorganic iron ;' higher plants. <A= %TPase proton pu-ps in the root cortical cells acidi>' the rhiAosphere8 ?hich helps to solu;iliAe as Fe.+ <FeIII=. The Fe.+ is then chelated ;' phenolic acids <"h=8 also secreted into the rhiAosphere ;' the roots. The chelated iron is carried to the root sur>ace ?here it is reduced ;' an FeIII reductase. The resulting Fe2+ <FeII= is i--ediatel' transported across the plas-a -e-;rane ;' an FeII transporter. *oth the FeIII reductase and the FeII are induced ;' iron de>icienc'. <B= Fe.+ is solu;iliAed ;' ph'tosiderophores <PS= secreted into the rhiAosphere ;' the root. The entire >errisiderophore <siderophore/iron co-ple(= is then taBen into the root cell ?here the iron is su;seCuentl' released.

include the formation of specialized transfer cells in the root epidermis, enhanced proton secretion into the soil surrounding the roots, and the release of strong ligands, such as ca>>eic acid ',igure 9.9(, by the roots. &imultaneously, there is an induction of reducing enzymes in the plasma membrane of the root epidermal cells. Acidification of the rhizosphere encourages chelation of the ,e1; with caffeic acid, which then moves to the root surface where the iron is reduced to ,e0; at the plasma membrane ',igure 9.DA(. Eeduction to ,e0; causes the ligand to release the iron, which is immediately taken up by the plant before it has the opportunity to form insoluble precipitates. A second strategy for iron uptake by organisms involves the synthesis and release by the organism of low-molecular-weight, iron-binding ligands called siderophores 'Gr. iron-bearers(. -ost of our knowledge of siderophores comes from studies with aerobic microorganisms 'bacteria, fungi, and algae(, where they were first discovered and have been studied

most extensively. -ore recently, however, it has been discovered that siderophores are also released by the roots of higher plants ',igure 9.I(. )nown as ph'tosiderophores, to distinguish them from ligands of microbial origin, these highly specific iron-binding ligands have thus far been found only in members of the family Gramineae, including the cereal grains. !hytosiderophores are synthesized and released by the plant only under conditions of iron stress, have a high affinity for ,e1;, and very effectively scavenge iron from the rhizosphere. The distinctive feature of the siderophore system is that the entire iron-phytosiderophore complex, or ferrisiderophore, is then reabsorbed into the roots ',igure 9.D=(. #nce inside the root, the iron is presumably reduced to ,e0; and released for use by the cell. The fate of the phytosiderophore is unknown. 7n microorganisms, siderophores may be chemically degraded and metabolized or, alternatively, the same 6# +## +## +##6 *60 #6
;

*60
;

+##6 #6 #6 6 +## ;*6 *60

;

Avenic acid (AA) u!inec acid ( A)

HH

+## H H
FI+U!E 4.9 Ph'tosiderophores. The structures o> t?o ph'tosiderophores released ;' the roots o> higher plants. Ferric iron >or-s coordinate ;onds ?ith the nitrogen and car;o('l groups. 4.4 Nutrient Functions and Deficiency Symptoms 4.

molecule may again be secreted by the cell in order to pick up more iron. The study of phytosiderophores is a relatively young field and, although substantial progress has been made in recent years, there is still much to be learned. 7t is not yet known, for example, how widespread the use of phytosiderophores is and the nature of the ferrisiderophore transport system has not been demonstrated in plants. #ne thing is clear" in those plants that use them, phytosiderophores are an important and effective strategy for supplying iron to the plant under conditions of iron stress.

4.4.D *O!ON %PPE%!S TO H%)E % !O#E IN "E## I)ISION %N E#ON+%TION %N "ONT!I*UTES TO THE

ST!U"TU!%# INTE+!ITY OF THE "E## ,%##

7n a$ueous solution, boron is present as boric acid, or 61=#1. At physiological p6 '<3(, it is found predominantly in the undissociated form, which is preferred for uptake by roots. :ith respect to its biochemical and physiological role, boron is perhaps the least understood of all the micronutrients. There is, for example, no solid evidence for involvement of boron with specific enzymes, either structurally or as an activator. 7ndeed, most of what we know about the role of boron is based entirely on studies of what happens to plants when boron is withheld. A substantial proportion of the total borate content of cells is found in the cell wall. This is apparently because borate has a propensity to form stable esters with cell wall saccharides that have ad acent hydroxyl groups. This so-called cis-diol configuration is characteristic of some common cell wall polysaccharides, such as mannose and its derivatives. Glucose, fructose, and galactose, on the other hand, do not have this configuration and so do not bind boron. The primary walls of boron-deficient cells exhibit marked structural abnormalities, suggesting that boron is re$uired for the structural integrity of the cell wall. #ther responses to boron deficiency point toward a role in cell division and elongation. #ne of the most rapid responses to boron deficiency, for example, is an inhibition of both cell division and elongation in primary and secondary roots. This gives the roots a stubby and bushy appearance. +ell division in the shoot apex and young leaves is also inhibited, followed by necrosis of the meristem. 7n addition, boron is known to stimulate pollen tube germination and elongation. 7t is not known how boron is involved in cell growth, but both hormone and nucleic acid metabolism have been implicated. 7nhibition of cell division and elongation is accompanied by an increased activity of enzymes that oxidize the hormone indole-1-acetic acid and a decrease in E*A content 'possibly through impaired synthesis of uracil, an E*A precursor(. 7n addition to the effects on shoot meristems noted above, common symptoms of boron deficiency include shortened internodes, giving the plant a bushy or rosette appearance, and enlarged stems, leading to the disorder known as AAstem crackBB in celery. 7n storage roots such as sugar beets, the disorder known as AAheart rotBB is due to the death of dividing cells in the growing region because of boron deficiency.

4.4.15 "OPPE! IS % NE"ESS%!Y "OF%"TO! FO! O0I %TI)E EN3Y$ES

7n well-aerated soils, copper is generally available to the plant as the divalent cupric ion, +u0;. +u0; readily forms a chelate with humic acids in the organic fraction

of the soil and may be involved in providing copper to the surface of the root. 7n wet soils with little oxygen, +u0; is readily reduced to the cuprous form, +u;, which is unstable. As a plant nutrient, copper seems to function primarily as a cofactor for a variety of oxidative enzymes. These include the photosynthetic electron carrier plastocyanin< cytochrome oxidase, which is the final oxidase enzyme in mitochondrial respiration< and ascorbic acid oxidase. The browning of freshly cut apple and potato surfaces is due to the activity of copper-containing pol'phenolo(idases 'or phenolase(. Supero(ide dis-utase <SO =, which detoxifies superoxide radicals '#5 0 (, is another important copper enzyme. +ommon disorders due to copper deficiency are generally stunted growth, distortion of young leaves and, particularly in citrus trees, a loss of young leaves referred to as AAsummer dieback.BB

4.4.11 3IN" IS %N %"TI)%TO! OF NU$E!OUS EN3Y$ES

Jinc is taken up by roots as the divalent cation Jn0;. Jinc is an activator of a large number of enzymes, including alcohol deh'drogenase <% H=, which catalyzes the reduction of acetaldehyde to ethanol< car;onic anh'drase <"%=, which catalyzes the hydration of carbon dioxide to bicarbonate< and, copper &#> which detoxifies #0 5. 6owever, there is general agreement that disorders associated with zinc deficiency reflect disturbances in the metabolism of the auxin hormone indole-1-acetic acid. Typically, zinc-deficient plants have shortened internodes and smaller leaves 'e.g., AAlittle leaf BB disorder of fruit trees(. The precise role of zinc in auxin metabolism remains obscure, but auxin levels in zinc-deficient plants are known to decline before the overt symptoms of zinc deficiency appear. ,urthermore, restoration of the zinc supply is followed by a

44 "hapter 4 6 Plants and Inorganic Nutrients

rapid increase in hormone level and then resumption of growth. Available evidence supports the view that zinc is re$uired for synthesis of the hormone precursor tryptophan.

4.4.12 $%N+%NESE IS %N EN3Y$E "OF%"TO! %S ,E## %S P%!T OF THE O0Y+EN/E)O#)IN+ "O$P#E0 IN THE "H#O!OP#%ST
-anganese is absorbed and transported within the plant mainly as the divalent cation -n0;. -anganese is re$uired as a cofactor for a number of enzymes, particularly decarboxylase and dehydrogenase enzymes, which play a critical role in the respiratory carbon cycle. 7nterestingly, manganese can often substitute for magnesium in reactions involving, for example, AT!. 6owever, the

best known and most studied function of manganese is in photosynthetic oxygen evolution '+hapter 8(. 7n the form of a -anganoprotein, manganese is part of the oxygen-evolving complex associated with photosystem 77, where it accumulates charges during the oxidation of water. -anganese deficiency can be widespread in some areas, depending on soil conditions, weather, and crop species. >eficiency is aggravated by low soil p6 '<I( and high organic content. -anganese deficiency is responsible for AAgray speckBB of cereal grains, a disorder characterized by the appearance of greenish-gray, oval-shaped spots on the basal regions of young leaves. 7t may cause extreme chlorosis between the leaf veins as well as discoloration and deformities in legume seeds.

4.4.1. $O#Y* ENU$ IS % 2EY "O$PONENT OF NIT!O+EN $ET%*O#IS$

Although molybdenum is a metal, its properties more closely resemble those of the nonmetals. 7n a$ueous solution it occurs mainly as the molybdate ion -oO05 9. -olybdenum re$uirements are among the lowest of all known micronutrients and appear to be primarily related to its role in nitrogen metabolism. Among the several enzymes found to re$uire molybdenum are dinitrogenase and nitrate reductase. The molybdenum re$uirement of a plant thus depends to some extent on the mode of nitrogen supply '+hapter //(. >initrogenase is the enzyme used by prokaryotes, including those in symbiotic association with higher plants, to reduce atmospheric nitrogen. *itrate reductase is found in roots and leaves where it catalyzes the reduction of nitrate to nitrite, a necessary first step in the incorporation of nitrogen into amino acids and other metabolites. 7n plants such as legumes, which depend on nitrogen fixation, molybdenum deficiency gives rise to symptoms of nitrogen deficiency. :hen nitrogen supplies are ade$uate, a deficiency of molybdenum shows up as a classic disorder known as AAwhiptailBB in which the young leaves are twisted and deformed. The same plants may exhibit interveinal chlorosis and necrosis along the veins of older leaves. Cike many of the micronutrients, molybdenum deficiency is highly species dependent?it is particularly widespread for legumes, members of the family =rassicaceae, and for maize. -olybdenum deficiency is aggravated in acid soils with a high content of iron precipitates, which strongly adsorb the molybdate ion.

4.4.14 "H#O!INE H%S % !O#E IN PHOTOSYNTHETI" O0Y+EN E)O#UTION %N "H%!+E *%#%N"E %"!OSS "E##U#%! $E$*!%NES

+hloride ion '+l5( is ubi$uitous in nature and highly soluble. 7t is thus rarely, if ever, deficient. >eficiencies normally can be shown only in very carefully controlled solution culture experiments. Along with manganese, chloride is re$uired for the oxygen-evolving reactions of photosynthesis '+hapter 8(. +l5 is a highly mobile anion with two principal functions" it is both a ma or counter-ion to diffusible cations, thus maintaining electrical neutrality across membranes, and one of the principal osmotically active solutes in the vacuole. +hloride ion also appears to be re$uired for cell division in both leaves and shoots. +hloride is readily taken up and most plants accumulate chloride ion far in excess of their minimal re$uirements. !lants deprived of chloride tend to exhibit reduced growth, wilting of the leaf tips, and a general chlorosis.

4.4.17 THE !O#E OF NI"2E# IS NOT "#E%!

*ickel is a relatively recent addition to the list of essential nutrient elements. *ickel is an abundant metallic element and is readily absorbed by roots. 7t is ubi$uitous in plant tissues, usually in the range of 4.4D to D.4mg )g5/ dry weight. #ne of the principal difficulties encountered in attempting to establish a role for nickel is its extremely low re$uirement. 7t has been estimated that the $uantity of nickel needed by a plant to complete one life cycle is approximately 044 ng, a re$uirement that can be met by the initial nickel content of the seed in most cases. 7n order to establish a nickel deficiency, it is necessary to undertake extensive purification of the nutrient salts and then grow several successive generations to seed in nickel-deficient solutions. The strongest evidence in favor of essential status for nickel is based on studies with legumes and cereal grains. 7n one such study of barley 'Hordeum vulgare(, the critical nickel
Summary 47

concentration for seed germination was found to be 24 ng g5/ of seed dry weight. =y growing plants for three generations in the absence of nickel, the nickel content of the seed could be reduced to 8.4 ng g 5/ dry weight. Germination of these seeds was less than /0 percent. :hen the plants were grown for the same number of generations in nutrient solution supplemented with 4.I - or /.4- nickel, seed germination was D8 and 2D percent, respectively. 7n other studies, nickel deficiencies have led to depressed seedling vigor, chlorosis, and necrotic lesions in leaves. The basis for a nickel re$uirement by plants is not clear, but it may be related to mobilization of nitrogen during seed germination. *ickel is known to be a component of two enzymes< urease and hydrogenase. Urease catalyzes the hydrolysis of urea into *61 and +#0 and is found widely through the plant kingdom. Urease from ack bean seeds ' anavalia ensiformis( was in fact the first protein to be crystallized by %. =. &umner

in /20I. #ne of the principal effects of nickel deficiency in soybean '!lycine max( is decreased urease activity in the leaves, although the metabolic significance of urease is not yet clear. ,ree urea is rarely, if ever, detected in plant tissue, but it is formed by the action of the enzyme arginase on arginine and its structural analog, canavanine '+hapter //(. +anavanine, a nonprotein amino acid, is abundant in the seeds of some plant groups, such as ack bean, but its concentration diminishes rapidly upon germination. Arginine is also abundant in seeds and both amino acids could function as stored nitrogen that is readily mobilized during seed germination. 7f this view should be proven valid, then urease, and thus nickel as well, would play an important role in the mobilization of nitrogen during germination and early seedling growth. A common form of mobile nitrogen in some legumes is a family of urea-based compounds known as ureides, such as allantoic acid or citrulline '+hapter //(. Ureides are formed in root nodules during nitrogen fixation and transported via the xylem throughout the host plant. Ureides are also formed in senescing leaves and transported out to the developing seeds for storage. The breakdown of ureides produces urea, which accumulates to toxic levels in *i-deficient plants. ,urthermore, the metabolism of purine bases 'adenine and guanine( in all plants also produces ureides. 7t seems reasonable to assume that most, if not all, plants have a re$uirement for urease and nickel. H'drogenase is another important enzyme in some nitrogen-fixing plants. 6ydrogenase is responsible for recovering hydrogen for use in the nitrogen-fixing process '+hapter //(. A deficiency of nickel leads to depressed levels of hydrogenase activity in the nodules of soybean, which in turn would be expected to depress the efficiency of nitrogen fixation.

4.7 TO0I"ITY OF $I"!ONUT!IENTS

As a group, the micronutrient elements are an excellent example of the dangers of excess. -ost have a rather narrow ade$uate range and become toxic at relatively low concentrations. "ritical to(icit' le@els, defined as the tissue concentration that gives a /4 percent reduction in dry matter, vary widely between the several micronutrients as well as between plant species. As noted earlier, critical concentrations for copper, boron, and zinc are on the order of 04, 8D, and 044 g g5/ dry weight, respectively. #n the other hand, critical toxicity levels for manganese vary from 044 g g5/ dry weight for corn, to I44 gg5/ for soybean, and D144 gg5/ for sunflower. Toxicity symptoms are often difficult to decipher because an excess of one nutrient may induce deficiencies of other nutrients. ,or example, the classic symptom of

manganese toxicity, which often occurs in waterlogged soils, is the appearance of brown spots due to deposition of -n#0 surrounded by chlorotic veins. =ut excess manganese may also induce deficiencies of iron, magnesium, and calcium. -anganese competes with both iron and magnesium for uptake and with magnesium for binding to enzymes. -anganese also inhibits calcium translocation into the shoot apex, causing a disorder known as AAcrinkle leaf.BB Thus the dominant symptoms of manganese toxicity may actually be the symptoms of iron, magnesium, andKor calcium deficiency. .xcess micronutrients typically inhibit root growth, not because the roots are more sensitive than shoots but because roots are the first organ to accumulate the nutrient. This is particularly true of both copper and zinc. +opper toxicity is of increasing concern in vineyards and orchards due to long-term use of copper-containing fungicides as well as urban and industrial pollution. Jinc toxicity can be a problem in acid soils or when sewage sludge is used to fertilize crops. 7n spite of the apparent toxicity of micronutrients, many plant species have developed the capacity to tolerate extraordinarily high concentrations. ,or example, most plants are severely in ured by nickel concentrations in excess of D g g5/ dry weight, but species of the genus Alyssum can tolerate levels in excess of /4 444 g g5/ dry weight.